In vitro effects of juvenile hormone on puffing in Chironomys salivary glands

Isolated salivary glands of Chironomus tentons prepupae were incubated in a synthetic culture medium containing 10^?4 M β-ecdysone and various amounts of C18-Cecropia juvenile hormone (JH). After 2 or 4 h their chromosomal puffing patterns were analyzed. Between 3.4 × 10^?7 and 3.4 × 10^?5 M, JH induces the ‘juvenile’ puff I-19-A₁. At higher concentrations (3.4 × 10^?4 M), a previous induction of this puff is reversed. The ecdysone-dependent puff I-18-C becomes progressively reduced at JH concentrations above 3.4 × 10^?6 M. Wounding of glands results in a transient, JH-independent induction and reduction, respectively, of puffs I-19-A₁ and I-18-C, and eventually, in the formation of a new puff (I-19-A₂); wounding generally diminishes the effects of JH on puffing. In the absence of ecdysone, puffs I-19-A₁ and I-19-A₂ develop spontaneously, and 3.4 × 10^?4 M JH no longer reverses puff formation in I-19-A₁. These results suggest an antagonistic interaction of JH and ecdysone with a cellular regulatory circuit involving cell membrane permeability changes.     

Impact of gender on insulin signaling pathway in lacrimal and salivary glands of rats

The structure and function of lacrimal and salivary glands present gender differences. Previous works have indicated a synergic action between insulin and androgens over lacrimal gland, and insulin-signaling pathways were recently described in lacrimal gland and salivary gland. Our present study investigates whether gender modulates the early steps of the insulin-signaling system in vivo. Eight-week-old male and female Wistar rats (n=8/group) were compared to evaluate insulin serum levels and insulin tolerance tests by radio-immunoassay and glucose oxidase method, respectively. To assess insulin receptor (IR), Shc, STAT-1, ERK, and Akt phosphorylation in response to insulin in lacrimal gland and salivary gland, tissues from female and male rats (n=5–8/group) were submitted to immunoprecipitation and immunoblotting or Western blotting protocol, and phosphorylation level was determined by densitometry. No difference was found in insulin serum levels or insulin tolerance tests comparing both groups. Nevertheless, lacrimal gland and salivary gland of female rats had a significantly lower insulin-induced IR phosphorylation compared with males. IR phosphorylation was not affected by the estrous cycle stage in either tissue. In addition, in females an apparent but not significant lower STAT and Akt phosphorylation in response to insulin was observed in the lacrimal gland, compared with males. Our findings suggest that alterations in insulin signal transduction may play a role in lacrimal gland and salivary gland gender differences.     

Solitary adrenal metastasis from salivary duct carcinoma of the parotid gland successfully treated by surgery: A case report

Rationale:Salivary duct carcinoma (SDC) is a rare and highly aggressive cancer with a poor prognosis. SDC demonstrates a potential for invasive growth with early regional and distant metastasis to organs, such as bone, lung, liver, and brain. Because, adrenal gland metastasis from SDC is rare, its treatment options are not well established. Herein, we report a case of SDC metastasis from the parotid gland to the adrenal gland, which was successfully treated by surgery.Patient concerns:The patient had an abnormal but painless lump on the right parotid gland. The size of the mass had increased over a period of 3 years. The patient underwent complete removal of the right parotid gland and radical neck dissection followed by adjuvant radiotherapy and chemotherapy. Two years later, a mass was identified in the left adrenal gland by computed tomography. As no local recurrence or metastasis to other organs was observed, the patient underwent adrenalectomy.Diagnoses:Metastasis of SDC in the adrenal gland was confirmed by histopathological examination of the adrenalectomized specimen.Interventions:After adrenalectomy, the patient was followed-up without adjuvant therapy.Outcomes:The patient was well and alive during the 13-month postoperative follow-up period without any complications.Lessons:Surgical resection of solitary metastatic lesion may show a survival benefit with metastatic SDC.     

Membrane association of importin α facilitates viral entry into salivary gland cells of vector insects

The importin α family belongs to the conserved nuclear transport pathway in eukaryotes. However, the biological functions of importin α in the plasma membrane are still elusive. Here, we report that importin α, as a plasma membrane–associated protein, is exploited by the rice stripe virus (RSV) to enter vector insect cells, especially salivary gland cells. When the expression of three importin α genes was simultaneously knocked down, few virions entered the salivary glands of the small brown planthopper, Laodelphax striatellus. Through hemocoel inoculation of virions, only importin α2 was found to efficiently regulate viral entry into insect salivary-gland cells. Importin α2 bound the nucleocapsid protein of RSV with a relatively high affinity through its importin β–binding (IBB) domain, with a dissociation constant K_D of 9.1 μM. Furthermore, importin α2 and its IBB domain showed a distinct distribution in the plasma membrane through binding to heparin in heparan sulfate proteoglycan. When the expression of importin α2 was knocked down in viruliferous planthoppers or in nonviruliferous planthoppers before they acquired virions, the viral transmission efficiency of the vector insects in terms of the viral amount and disease incidence in rice was dramatically decreased. These findings not only reveal the specific function of the importin α family in the plasma membrane utilized by viruses, but also provide a promising target gene in vector insects for manipulation to efficiently control outbreaks of rice stripe disease.

The importin α family belongs to the conserved importin α/β nuclear transport pathway in eukaryotes (1–3). It is well known that the importin α family plays an indispensable role in transporting proteins from the cytoplasm to the nucleus, with diverse functions in gene regulation, cell physiology, and differentiation (1, 4, 5). In addition to nucleocytoplasmic transport, some members of the importin α family localize to the plasma membrane with palmitoylation modification or through binding to heparin in heparan sulfate proteoglycan (HSPG) (6–8). Increased importin α levels in the plasma membrane are concomitant with decreased importin α levels in the cytoplasm, which affect the nuclear import of cargos and regulates intracellular scaling (7). However, the function of the importin α family in the plasma membrane is still elusive.Many plant viruses are transmitted by vector insects in a persistent, circulative mode, which is characterized by systemic invasion of diverse tissues prior to entering salivary glands and release in saliva (9–13). The salivary glands are the last barriers for viral transmission to overcome (14–18). Unfortunately, the molecular mechanisms underlying viral entry into salivary-gland cells are not well known. The rice stripe virus (RSV) is a typical persistent, circulative plant virus and is capable of proliferating in the midgut epithelial cells and of being efficiently transmitted by the vector insect small brown planthopper (Laodelphax striatellus) (19). This virus causes one of the most destructive rice stripe diseases, showing an incidence of up to 80% and causing yield losses of 30 to 40% in the rice fields of Asian countries (20). RSV is a nonenveloped negative-strand RNA virus of the Tenuivirus genus (21, 22). The genome of RSV contains four single-stranded RNA segments, each of which is encapsidated by a viral nucleocapsid protein (NP). In addition to the NP, RSV encodes an RNA-dependent RNA polymerase and five nonstructural proteins (NS2, NSvc2, NS3, SP, and NSvc4) (23–25).In our recent work, we found that three importin α proteins, importin α1 (GenBank registration number {"type":"entrez-nucleotide","attrs":{"text":"MT036051","term_id":"2078901399","term_text":"MT036051"}}MT036051), α2 ({"type":"entrez-nucleotide","attrs":{"text":"MT036050","term_id":"2078901365","term_text":"MT036050"}}MT036050), and α3 ({"type":"entrez-nucleotide","attrs":{"text":"MT036052","term_id":"2078901435","term_text":"MT036052"}}MT036052), of the small brown planthopper participate in the nuclear entry of RSV through direct interactions with RSV NPs, triggering an antiviral caspase-dependent apoptotic reaction (26). Knockdown of the expression of all the three importin α genes retarded the nuclear entry of RSV and led to an increase in viral load in planthoppers (26). However, we did not determine the influence on viral transmission. In the present study, we surprisingly found that one of the importin α proteins, importin α2, is associated with the plasma membrane and efficiently facilitates viral entry into insect salivary glands and subsequent viral transmission.     

斑须按蚊唾液腺中疟原虫配子体激活因子的活性动态

目的　探讨斑须按蚊生长、发育过程唾液腺中疟原虫配子体激活因子的活性动态。方法　应用体外雄配子体出丝分析方法 ,检测雌性斑须按蚊羽化后、吸血及产卵前后唾液腺中疟原虫配子体激活因子对柏氏疟原虫雄配子体出丝诱导活性的变化。结果　羽化后未吸血组斑须按蚊唾液腺配子体激活因子活性与按蚊生长、发育呈同步变化 ;吸血后未产卵组按蚊唾液腺抽提物的配子体激活因子活性在吸血后显著下降 ,吸血后第 8天恢复到吸血前水平 ;而吸血后产卵组按蚊唾液腺抽提物的配子体激活因子活性在吸血后第 4天恢复到吸血前水平。结论　吸血后斑须按蚊唾液腺配子体激活因子的消长与按蚊产卵相关。     

The diagnostic utility of labial salivary gland biopsy in IgG4-related disease

Objective: For the definitive diagnosis of IgG4-related disease (IgG4-RD), biopsies of local lesions are recommended so as to exclude other diseases, including lymphoma and cancer. However, performing biopsies of underlying organs is technically difficult. In this study, we examined the diagnostic utility of labial salivary gland (LSG) biopsy as a less invasive procedure.

Methods: Sixty-six patients with suspected IgG4-RD by clinical findings or high serum IgG4 underwent LSG biopsy. We examined the relationship between the number of IgG4-positive plasma cells in LSG and clinical findings.

Results: The final diagnosis was 45 patients with IgG4-RD, 12 with Sjögren’s syndrome, four with suspected Sjögren’s syndrome, three with malignant lymphoma, one with systemic lupus erythematosus, and one with Warthin’s tumor. The sensitivity, specificity, and accuracy of LSG biopsy were 55.6%, 100.0%, and 70.0%, respectively. Forty-five IgG4-RD patients were divided into two groups: 1) 25 with lesions of salivary glands (IgG4-RD S+) and 2) 20 without these lesions (IgG4-RD S?). Seventeen of 25 (68.0%) IgG4-RD S?+?and 8 of 20 (40.0%) IgG4-RD S???patients were positive for LSG biopsy. In the IgG4-RD S???patients, the mean number of affected organs and serum IgG4 in the positive cases for LSG biopsy were significantly higher than in the negative cases.

Conclusion: A solo LSG biopsy is insufficient for the diagnosis of IgG4-RD because of its low sensitivity. However, LSG biopsy combined with clinical findings, including serum IgG4 and number of affected organs, may contribute towards a diagnosis of IgG4-RD patients with affected underlying organs.     

Most cases of primary salivary mucosa-associated lymphoid tissue lymphoma are associated either with Sjoegren syndrome or hepatitis C virus infection

Salivary gland mucosa-associated lymphoid tissue (MALT) lymphomas (SGML) are rare, as are data concerning their behaviour. We analysed clinical features at presentation, particularly the association with Sjoegren syndrome (SS) and hepatitis C virus (HCV) infection, and outcome in 33 cases of SGML diagnosed between March 1985 and April 2003. There were five males and 28 females, with a median age of 61 years. At presentation, 12/33 (36%) had multiple salivary glands or mucosal involvement and four had bone marrow infiltration. Ann Arbor stage was IE in 15 (46%), IIE in four (12%) and IV in 14 patients (42%). Fifteen patients had a history of SS (46%), two of other autoimmune diseases, seven of HCV infection. No case had both SS and HCV. Of the 29 treated patients, 17 received surgery or local radiotherapy; 69% achieved complete remission. Histological transformation occurred in four (12%). Five patients died (three of lymphoma, two of unrelated causes). The 5 year-overall survival (OS), cause-specific survival and progression-free survival was 85 +/- 8%, 94 +/- 6% and 65 +/- 10% respectively. Overall, the disease course was indolent, despite the advanced stage at diagnosis, and local therapy often appeared to be adequate. The only prognostic factors influencing OS were histological transformation and age. The close association of SGML with either autoimmune diseases or HCV infection in our series (73%) confirms their possible role in the pathogenesis of these lymphomas.     

Extracts of mosquito salivary gland inhibit tumour necrosis factor alpha release from mast cells

Extracts of salivary glands of the yellow fever mosquito Aedes aegypti inhibit tumour cell-stimulated release of tumour necrosis factor alpha (TNFα) from rat mast cells, but do not inhibit antigen-induced histamine secretion. This inhibitory activity for TNFα is found in salivary glands of female but not in male mosquitoes. This inhibition is not mediated by bacterial contamination (LPS), by calcitonin gene related peptide (CGRP), nerve growth factor (NGF), epidermal growth factor (EGF) or transforming growth factor β (TGFβ). The factor(s) has a molecular weight > 10 kDa and is neutralized by boiling for 10 min or heating at 56°C for 30 min. The modulation of this proinflammatory mediator, TNFα, produced by mast cells in sites of blood feeding may facilitate completion of the blood meal, and as reported for certain vector-transmitted parasites, may enhance infectivity.     

A serological study of intrafamilial spread from patients with sporadic hepatitis E virus infection

Summary. Intrafamilial transmission is rare in epidemic hepatitis E; its frequency in sporadic hepatitis E is not known. We followed up 86 household contacts (age range 4–75 years, mean ± SD 32.4 ± 15.8; 49 males), who were family members of patients with acute sporadic hepatitis E. Of the 86 contacts, 68 (79%) tested negative for IgG anti-hepatitis E virus antibodies. Four (4.7%) had IgM anti-hepatitis E virus antibodies at the time of diagnosis of hepatitis E in the index case; two of these contacts possibly had hepatitis E virus infection acquired simultaneously with that in the index case, and two could have had intrafamilial transmission. None developed serological evidence of hepatitis E virus infection over a period of 49 ± 18 days after the diagnosis of index case, although a majority lacked IgG antibodies to hepatitis E virus and were likely to be susceptible. This suggests that person-to-person transmission is uncommon in sporadic hepatitis E.     

Isolation and characterization of salivary antigens from the female tick, Dermacentor andersoni

The salivary glands of ixodid ticks are complex organs which are known to contain the antigens responsible for tick resistance in animals. We have identified a large number of proteins from salivary gland extracts (SGE), at least some of which are immunologically recognized by tick resistant animals and which are therefore presumed to be secreted salivary components. During the 6 to 10 day feeding process, a number of these antigens alter in concentration according to individual kinetics, and some of these changes correlate with the kinetics of skin test reactivity of SGE obtained at different times throughout the feeding period. By use of immunoaffinity chromatography we have isolated large quantities of many of the salivary antigens (SGA) contained in SGE, and found that they contain several esterase activities. SGA stimulates both immediate and delayed skin reactions in tick resistant guinea-pigs, and these reactions are about 200-fold more intense, per unit protein, than those elicited by SGE. The skin reactions to SGA are basophil-mediated and have many features in common with the cutaneous basophil hypersensitivity reactions of tick resistant animals. The demonstrated antigenic complexity of the glands may have profound implications for attempts to develop anti-tick vaccines, as it may eventually be found that candidate vaccines will have to incorporate more than one tick antigen in order to be effective.     

The clinical characteristics of patients with IgG4-related disease with infiltration of the labial salivary gland by IgG4-positive cells

Abstract

Objectives. Mikulicz's disease (MD) is an immunoglobulin (Ig) G4-related disease with systemic symptoms. Submandibular gland (SMG) biopsy is recommended for patients with possible IgG4-related MD for accurate differential diagnosis; however, it is difficult for certain patients to undergo this procedure. In contrast, labial salivary gland (LSG) biopsy is more convenient. Here we present an analysis of patients with IgG4-related MD whose LSG specimens were infiltrated with abundant IgG4-positive plasma cells.

Methods. Sixteen patients diagnosed with IgG4-related MD underwent simultaneous SMG and LSG biopsies. We evaluated patients’ serological and ¹⁸F-fluoro-2-deoxyglucose-positron emission tomography/computed tomography (¹⁸F-FDG-PET/CT) and grouped them as LSG+ (LSG specimens with > 40% IgG4-positive plasma cells/IgG-positive plasma cells, 11 patients) or LSG? (LSG specimens with < 40% IgG4-positive plasma cells/IgG-positive plasma cells, 6 patients).

Results. There were not significant differences in serum IgG and IgG4 levels between the two groups; however, serum concentrations of soluble interleuikin-2 receptor (sIL-2R) were significantly higher in the LSG+ group. All patients with increased ¹⁸F-FDG uptake in their parotid glands were a part of the LSG+ group.

Conclusions. When a SMG biopsy is not possible, the serum concentration of sIL-2R and ¹⁸F-FDG-PET/CT findings may predict whether LSG biopsy will facilitate the diagnosis of IgG4-related MD.     

胎盘滋养层细胞的乙型肝炎病毒感染与宫内感染机制

目的：检测HBV对胎盘、胎肝和滋养层细胞的感染情况，探讨HBV的宫内感染机制。方法：研究对象包括20例孕妇胎盘组织、6例引产胎儿胎肝组织及体外培养的胎盘滋养层细胞。ELISA法检测孕妇外周血、胎儿脐血和6个月婴儿外周血HBV标志物；荧光定量PCR法检测血清和滋养层细胞中的HBV DNA；免疫组织化学法和免疫荧光法检测胎盘、胎肝组织及滋养层细胞中HBV标志物的表达；末端脱氧核糖核酸转移酶介导的缺口标记法（TUNEL）检测胎盘和滋养层细胞凋亡情况。结果：孕妇血清HBV DNA水平与胎儿脐血HBV DNA水平相关，脐血HBV DNA阳性者其母血HBV DNA〉1.0×10^7拷贝／mL；6例胎盘组织和3例引产胎儿胎肝组织中可见HBsAg免疫组织化学染色阳性细胞，其中1例胎肝组织中发现HBcAg阳性细胞；体外培养滋养层细胞与HBV DNA阳性血清共孵育后，可检测到HBsAg的表达，亦可检测到HBV DNA。体内和体外实验均检测到HBV感染后滋养层细胞凋亡呈增加趋势，且胎盘细胞的凋亡与脐血HBV DNA水平相关。体外实验结果显示，随感染时间的延长，滋养层细胞凋亡呈增加趋势。6个月后，12例新生儿有1例血清HBsAg、HBeAg和抗-HBc阳性，6例抗-HBs阳性。结论：HBV宫内感染的机制可能是通过HBV感染胎盘屏障而使胎儿发生HBV宫内感染。HBV在胎儿组织器官内的定位和复制可能是新生儿发生慢性HBV感染的重要因素。滋养层细胞凋亡可能是胎盘屏障阻断HBV宫内传播的一种保护性机制。     

Hepatitis C virus infection of salivary gland epithelial cells: Lack of evidence

Background: Hepatitis C virus genome (HCV-RNA) has been detected in whole salivary gland tissue of chronically infected patients. However, contamination of the tissue by plasma or blood cells was not excluded by the previous reports.Aims: To assess whether HCV infects the salivary gland epithelial cells in patients with chronic HCV liver disease.Methods: Twenty unselected patients with chronic active hepatitis (11 cases) or active cirrhosis (nine cases) were examined. Serum and saliva samples were obtained from all patients, 12 of whom (seven, chronic active hepatitis; five, active cirrhosis) underwent salivary gland biopsy. PCR for HCV-RNA was performed on RNA extracted from serum, saliva and salivary gland epithelial cells collected by isokinetic gradient separation after trypsin digestion of whole salivary gland tissue. Saliva samples were also examined for the presence of secretory IgA anti-HCV by gel chromatography and ELISA testing.Results: HCV-RNA was detected in all sera with titers ranging from 5.42×10⁵ genome equivalents/ml to 123.2×10⁵ genome equivalents/ml. Thirteen patients were infected with genotype 1b, four patients had genotype 1a, two patients had genotype 2a and one patient was unclassifiable. Low titer HCV-RNA (<2×10⁵ genome equivalents/ml) was detected in saliva samples (15%) from highly viremic patients infected with 1b genotype. RNA extracted from salivary gland epithelial cells consistently tested negative for HCV-RNA. In addition, all saliva specimens tested negative for secretory-IgA (S-IgA) anti-HCV, even after a 10-fold concentration of the samples.Conclusions: There was no evidence that HCV infects the salivary gland epithelial cells in our viremic patients with HCV chronic liver disease. Low level HCV-RNA in saliva is most probably due to virus spillover from blood.     

Relationship of hepatitis B virus infection of placental barrier and hepatitis B virus intra-uterine transmission mechanism

AIM： To explore the mechanism of intra-uterine transmission, the HBV infection status of placental tissue and in vitro cultured placental trophoblastic cells was tested through in vivo and in vitro experiments. METHODS： A variety of methods, such as ELISA, RT- PCR, IHC staining and immunofluorescent staining were employed to test the HBV marker positive pregnant women＇s placenta and in vitro cultured placental trophoblastic cells. RESULTS： The HBV DNA levels in pregnant women＇s serum and fetal cord blood were correlated. For those cord blood samples positive for HBV DNA, their maternal blood levels of HBV DNA were at a high level. The HBsAg IHC staining positive cells could be seen in the placental tissues and the presence of HBV DNA detected. After coincubating the trophoblastic cells and HBV DNA positive serum in vitro, the expressions of both HBsAg and HBV DNA could be detected. CONCLUSION： The mechanism of HBV intra-uterine infection may be due to that HBV breaches the placental barrier and infects the fetus.     

An overview of occult hepatitis B virus infection

Occult hepatitis B virus (HBV) infection (OBI), alternatively defined as occult hepatitis B (OHB), is a challenging clinical entity. It is recognized by two main characteristics: absence of HBsAg, and low viral replication. The previous two decades have witnessed a remarkable progress in our understanding of OBI and its clinical implications. Appropriate diagnostic techniques must be adopted. Sensitive HBV DNA amplification assay is the gold standard assay for detection of OBI. Viral as well as host factors...     

Effect of salivary gland extracts from the tick,Boophilus microplus,on leucocytes from Brahman and Hereford cattle

The effect of salivary gland extract (SGE) from Boophilus microplus on peripheral blood lymphocytes, neutrophils and monocytes from Brahman (Bos indicus) and Hereford (Bos taurus) cattle was investigated. SGE (8 micro g) significantly inhibited the proliferation response of lymphocytes to concanavalin A from both Brahman and Hereford cattle by 89% and 41%, respectively. The difference in inhibition between the two breeds was highly significant (P < 0.01), whilst at 1 micro g of SGE, significant inhibition of lymphocytes occurred only in Hereford cattle (34%). Flow cytometric analysis of monocytes and neutrophils showed that SGE (40 micro g) significantly reduced both the proportion of cells actively phagocytosing Escherichia coli labelled with fluorescein isothiocyanate (E. coli-FITC) and the uptake of E. coli-FITC in Brahman cattle. However, in Hereford cattle, a significant depression in uptake was only observed in neutrophils. The proportion of monocytes and neutrophils with oxidative activity was significantly suppressed in the presence of SGE in both breeds of cattle. These results indicate that peripheral blood leucocytes from different breeds of cattle respond differently to SGE.     

Next-generation point-of-care testing in pediatric human immunodeficiency virus infection facilitates diagnosis and monitoring of treatment

Point-of-care (PoC) testing facilitates early infant diagnosis (EID) and treatment initiation, which improves outcome. We present a field evaluation of a new PoC test (Cepheid Xpert® HIV-1 Qual XC RUO) to determine whether this test improves EID and assists the management of children living with human immunodeficiency virus (HIV) infection.We compared 2 PoC tests with the standard-of-care (SoC) test used to detect HIV infection from dry blood spots in newborn infants at high risk of in utero infection. We also evaluated the ability of the PoC tests to detect HIV total nucleic acid (TNA) in children living with HIV infection who had maintained undetectable plasma viremia following very early combination antiretroviral therapy (cART) initiation.Qualitative (Qual) detection of HIV using the Xpert® HIV-1 Qual XC RUO (“RUO”) and Xpert® HIV-1 Qual (“Qual”) PoC tests was compared in 224 infants with the SoC DBS Roche COBAS® HIV-1/HIV-2 qualitative test. The same 2 PoC tests were also evaluated in 35 older children who had initiated cART before 21 days of age and maintained undetectable plasma viremia for a mean of 25 months.No discrepancies were observed in detection of HIV infection via the 2 PoC tests or the SoC test in the 224 neonates studied, but only 95% of the SoC test results were generated compared with 100% of the PoC test results (P = .0009). The cycle threshold values for the research use only (RUO) assay were the lowest of the 3 assays (P < .0001 in each case). In 6 of the 35 early-treated aviremic children, HIV TNA was detected by RUO but not Qual.The RUO assay outperforms Qual in detecting HIV-1 infection. RUO would therefore potentially improve EID and assist in identifying cART-adherent early-treated children with the lowest HIV TNA levels and the highest HIV cure potential.