Ultimobranchial gland of a freshwater teleost,Heteropneustes fossilis,in response to cadmium treatment

Heteropneustes fossilis were subjected to 288 mg/L (0.8 of 96 h LC₅₀) and 72 mg/L (0.2 of 96 h LC₅₀) of cadmium chloride for short‐term and long‐term experiments, respectively. After sacrificing the fish, the blood was collected on 24, 48, 72, and 96 h in short‐term and after 7, 14, 21, and 28 days in long‐term experiment and analyzed for plasma calcium levels. Also, ultimobranchial glands were fixed on these intervals. The plasma calcium levels of short‐term cadmium‐exposed fish remain unchanged after 24 h. The levels exhibit a progressive decrease from 48 h onwards. The fish exposed to cadmium for 7 days exhibit a decrease in the plasma calcium level. Thereafter, the levels progressively decrease till the end of the experiment (28 days). Up to 72 h exposure of the fish to cadmium, the ultimobranchial gland exhibits no histological change. After 96 h, a decrease in the staining response of the cytoplasm of ultimobranchial cells has been noticed. The nuclear volume of these cells records a slight decrease. Up to 14 days of cadmium exposure, there is no change in the histological structure of ultimobranchial gland. After 21 days following the exposure, the ultimobranchial cells exhibit a slight decrease in the staining response of the cytoplasm and the nuclear volume of these cells records a decrease. Following 28 days cadmium exposure the nuclear volume exhibits a further decrease, and degeneration and vacuolization sets in. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009.     

Histological alterations in the ultimobranchial gland of teleost Heteropneustes fossilis in response to chlorpyrifos treatment

In this study, an experiment was performed on Heteropneustes fossilis for short-term (1.76 mg/L chlorpyrifos, i.e., 0.8 of 96-h LC50) and long-term (0.44 mg/L chlorpyrifos, i.e., 0.2 of 96-h LC50) exposure. The fish were sacrificed after 24, 48, 72 and 96 h in the short-term experiment and after 7, 14, 21 and 28 days in the long-term experiment. On these intervals, blood was collected and analysis of serum calcium was done. Ultimobranchial glands were also fixed for histological study. The serum calcium levels of H. fossilis exhibit a decline after 24 h following exposure to chlorpyrifos. This decrease continues until the end of the experiment (96 h). The serum calcium levels of chronically exposed fish exhibit a decrease on day 7. Thereafter, the levels continue to fall progressively until the end of the experiment (28 days). The ultimobranchial gland of chlorpyrifos treated fish exhibits no histological change up to 48 h. After 72 h, there is a decrease in the staining response of cytoplasm of the ultimobranchial cells. The nuclear volume of these cells is slightly decreased. After 96 h following chlorpyrifos exposure, these changes become exaggerated. In chlorpyrifos-treated fish there is no change in the histological structure of the ultimobranchial gland up to 14 days. After 21 days, the cytoplasm of ultimobranchial cells stain feebly and the nuclear volume of these cells exhibits a decrease. Following 28 days treatment, the nuclear volume of these cells records a further decrease and the gland depicts vacuolization and degeneration at certain areas.     

Histological alterations in the prolactin cells of a teleost,Heteropneustes fossilis,after exposure to cypermethrin

Freshwater fish Heteropneustes fossilis (H. fossilis) were subjected to 5.76 μg/L (80% of 96 h LC₅₀) and 1.44 μg/L (20% of 96 h LC₅₀) of cypermethrin for short‐term (96 h) and long‐term (28 days) duration, respectively. Plasma calcium of H. fossilis exposed for short term (96 h) to cypermethrin exhibited no change at 24 h. The levels indicate a decrease in plasma calcium at 48 h. This response persists till the close of experiment (96 h). No change has been noticed throughout the experiment in the histological structure and nuclear volume of prolactin cells of short‐term cypermethrin treated fish. Long‐term exposure of cypermethrin to fish provoked hypocalcemia. The prolactin cells remain unchanged till 7 days following cypermethrin treatment. After 14 days, the nuclear volume exhibits an increase and the cells exhibit degranulation. These changes increase progressively 21 days onwards. Also, few degenerating cells are discerned after 28 days. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2011.     

Assessment of the acute toxicity of chlorpyrifos and cypermethrin to Heteropneustes fossilis and their impact on acetylcholinesterase activity

In the present study, the acute toxicity of chlorpyrifos (an organophosphate, OP) and cypermethrin (a pyrethroid) pesticides was estimated for 96?h in Heteropneustes fossilis. The LC₅₀ for chlorpyrifos (CPF) and cypermethrin was found to be 1.90?mg/L and 0.085?mg/L, respectively. The acetylcholinesterase (AChE, EC 3.1.1.7) activity in Heteropneustes fossilis exposed to both the insecticides was assayed in brain, muscle and gills. In general, tissue specific as well as dose-dependent decrease in the AChE activity was exhibited by both pesticides. In response to the increasing concentrations of chlorpyrifos and cypermethrin as well, a significant decrease in the activity of AChE was found in brain while muscle and gills exhibited lesser inhibition. Thus, the brain was the main target organ for both insecticides, followed by muscle and gills. Between the two pesticides chlorpyrifos acted as more potent AChE inhibitor than cypermethrin since more intense changes in behavioral pattern was observed with the chlorpyrifos. These changes indicate that the effects of these pesticides are at neural as well as neuromuscular level.     

Molluscicidal activity of cardiac glycosides from Nerium indicum against Pomacea canaliculata and its implications for the mechanisms of toxicity

Cardiac glycosides from fresh leaves of Nerium indicum were evaluated for its molluscicidal activity against Pomacea canaliculata (golden apple snail: GAS) under laboratory conditions. The results showed that LC₅₀ value of cardiac glycosides against GAS was time dependent and the LC₅₀ value at 96 h was as low as 3.71 mg/L, which was comparable with that of metaldehyde at 72 h (3.88 mg/L). These results indicate that cardiac glycosides could be an effective molluscicide against GAS. The toxicological mechanism of cardiac glucosides on GAS was also evaluated through changes of selected biochemical parameters, including cholinesterase (ChE) and esterase (EST) activities, glycogen and protein contents in hepatopancreas tissues of GAS. Exposure to sublethal concentrations of cardiac glycosides, GAS showed lower activities of EST isozyme in the later stages of the exposure period as well as drastically decreased glycogen content, although total protein content was not affected at the end of 24 and 48 h followed by a significant depletion at the end of 72 and 96 h. The initial increase followed by a decline of ChE activity was also observed during the experiment. These results suggest that cardiac glycosides seriously impair normal physiological metabolism, resulting in fatal alterations in major biochemical constituents of hepatopancreas tissues of P. canaliculata.     

Exposure and recovery response of isomers of HCH,metabolites of DDT and estradiol-17β in the female catfish,Heteropneustes fossilis

Effects of 40 days of exposure and 20 days of recovery response at sublethal concentration of technical grades of gamma isomer of hexachlorocyclohexane (γ-HCH, 0.025 ppm, 99.8%) and dichlorodiphenyltrichloroethane (DDT, 5.0 ppm) in tissue (liver, brain and ovary) bioconcentrations, gonadosomatic index (GSI) and plasma levels of estradiol-17β (E2) have been estimated during prespawning phase in the catfish Heteropneustes fossilis (Bloch). The results indicated that the tissue bioconcentrations of both HCHs (HCH isomers) and DDTs (metabolites of DDT) in liver, brain and ovary were in preferential order (liver > brain > ovary). The GSI and plasma levels of E2 were declined in response to exposure of γ-HCH and DDT. On withdrawal of exposure of pesticide there was recovery of HCHs in exposed fish for all tissues studied, whereas DDTs exposed fish showed recovery only in liver. Recovery of E2 production was also recorded in γ-HCH exposed fish whereas very little recorded in DDT exposed fish. It is suggested that HCHs and DDTs have preferential order (liver > brain > ovary) of their tissue bioconcentrations and HCH/DDT-withdrawal-dependent recovery during studied phase.     

肾上腺腺瘤样瘤临床病理观察

目的探讨肾上腺腺瘤样瘤的临床病理特征。方法报道1例64岁男性肾上腺腺瘤样瘤,包括其临床、影像及病理学特征,并复习相关文献。结果肾上腺腺瘤样瘤多发生于左侧,影像学无特征性表现;组织病理学特征,包括浸润周围组织、印戒样细胞,可能考虑侵袭性肿瘤,免疫表型calretinin、CK5/6、CK、EMA、Vim阳性。结论肾上腺腺瘤样瘤是一种罕见的良性病变,仅靠CT和磁共振成像(MRI)很难与肾上腺常见肿瘤相鉴别,根据病理组织形态学特点并结合免疫组化染色,可以明确诊断。     

Two volatiles from the venom gland of the Samsum ant, Pachycondyla sennaarensis

Allergy and anaphylactic reactions after the stings of the Samsum ant Pachycondyla sennaarensis (Mayr, 1862) (Hymenoptera: Formicidae) have been reported from several countries along the Persian Gulf coast, but no analysis has been yet carried out on the ant's venom gland secretions. This study is focused on the identification of volatiles from the venom gland of Pachycondyla sennaarensis using gas chromatography-mass spectrometry, which showed the presence of The main volatile components of the venom gland were phenol-2,4-bis(1,1 dimethylethyl) and trimethyl pyrazine. This is the first record of the occurrence of phenol-2,4-bis(1,1 dimethylethyl) in insects. The venom gland secretions of Pachycondyla species are known contain a variety of volatiles, making the members of this genus distinctive among the ponerine ants.     

Purification and molecular cloning of SE-cephalotoxin, a novel proteinaceous toxin from the posterior salivary gland of cuttlefish Sepia esculenta

Cephalopods contain toxins in their salivary glands, presumably to paralyze prey animals such as crabs and bivalves. Proteinaceous toxins (called cephalotoxins) with crab lethality have previously been purified from three species of octopodiform cephalopods (octopuses) but their detailed properties and primary structures have remained unknown. In this study, salivary glands of six species of decapodiform cephalopods were newly found to be toxic; three species of cuttlefish were lethal only to crabs and three species of squid to both mice and crabs. A proteinaceous toxin (named SE-cephalotoxin) in the salivary gland of cuttlefish Sepia esculenta was soluble only in high-salt solvents. This unique solubility enabled us to purify SE-cephalotoxin by gel filtration HPLC and hydroxyapatite HPLC. SE-cephalotoxin was shown to be a 100kDa monomeric glycoprotein with an LD(50) (against crabs) of 2mug/kg. Based on the determined partial amino acid sequence, a full-length cDNA (3402bp) coding for SE-cephalotoxin was cloned by RT-PCR and RACE. The SE-cephalotoxin precursor protein (1052 amino acid residues) is composed of a signal peptide (region 1-21), propeptide (region 22-29) and mature protein (region 30-1052). A database search failed to find any proteins sharing homology with SE-cephalotoxin.     

The effect of K channel openers on submucosal gland function and epithelial transport of the ferret trachea, in vitro

The effects of three K⁺ channel openers on lysozyme output from submucosal gland serous cells and epithelial albumin transport following maintained submaximal stimulation by the secretagogues methacholine and phenylephrine were examined in the ferret trachea in vitro preparation. The K⁺ channel openers Ro 31-6930, 2-(6-cyano-2,2-dimethyl-2H-1-benzopyran-4-yl)-pyridine 1-oxide (10 nM-10 μM), levcromakalim, BRL38227 (10 nM-10 μM) and pinacidil (100 nM-10 μM) produced a concentration dependent inhibition of (20 μM) methacholine-induced lysozyme output, with pD₂ values of 7.64, 7.72 and 7.28 respectively. Ro 31-6930 (10 nM-10 μM), levcromakalim (10 nM-10 μM) and pinacidil (1 nM-10 μM) also produced a concentration dependent inhibition of (100 μM) phenylephrine-induced lysozyme output, with pD₂ values of 7.64, 6.55 and 9.16 respectively. Furthermore, glibenclamide (1 μM) produced a modest attenuation of the K⁺ channel opener effects on secretagogue-induced lysozyme output. All three K⁺ channel openers failed to produce any significant change in either methacholine or phenylephrine-induced albumin outputs. The K⁺ channel openers exerted marked effects on airway secretion processes, suggesting that these compounds may have an antisecretory effect. The relevance of the use of the K⁺ channel openers in airway disease remains to be determined.     

IgG4相关疾病累及腮腺1例并文献复习

目的 探讨IgG₄相关疾病(IgG₄-related diseases,IgG₄-RD)累及腮腺的临床表现、诊断标准、治疗方案以及细针穿刺在疾病诊断中的应用,减少临床误诊漏诊。方法 回顾分析华北理工大学附属医院2020年11月收治的1例累及腮腺的IgG₄-RD病人的病例资料,通过分析临床特点、血清学指标、影像学表现及术后病理学特征等,并复习相关文献进行分析总结。结果该66岁男性病人以右侧腮腺无痛性肿物为主要表现,行抗感染治疗后肿物无明显变化,先后两次行细针穿刺均未明确诊断,行手术切除后病理结果确诊为累及腮腺的IgG₄-RD,结合病人术前检查及术后病理结果,考虑病变处于非活动期,暂未给予药物治疗,密切随访,至今未见复发。结论 IgG₄相关疾病为临床罕见疾病,缺乏典型的临床表现及特异的影像学特征,术前多不能明确诊断,病理学检查在疾病的诊断中具有重要意义,而细针穿刺在疾病的诊断中价值有限,需引起临床医生的重视。     

Primary structure of echotoxin 2, an actinoporin-like hemolytic toxin from the salivary gland of the marine gastropod Monoplex echo.

Echotoxins are 25 kDa proteins with both hemolytic and lethal activities, previously purified from the salivary gland of the marine gastropod Monoplex echo. In this study, a cDNA encoding echotoxin 2 was cloned by RT-PCR, 3'-RACE and 5'-RACE, based on its partial amino acid sequence. The full-length echotoxin 2 cDNA (1000 bp) obtained contains an open reading frame (825 bp) coding for a precursor protein of 274 amino acid residues. Mature echotoxin 2 composed of 226 amino acid residues is assumed to be produced by post-translational removal of N-terminal 23 residues (predicted as a signal peptide) and C-terminal 25 residues from the precursor protein. Very interestingly, a homology search revealed that echotoxin 2 is analogous to actinoporins, 20 kDa pore-forming hemolysins reported from various sea anemones. In addition to the similarities in biological activity, molecular size and basicity between echotoxin 2 and actinoporins, two prominent structural features, an N-terminal amphiphilic alpha-helix and an aromatic patch comprising Trp and Tyr residues, both of which are important for the pore-forming activity of actinoporins, are also recognized in echotoxin 2. However, echotoxin 2 is distinguishable from actinoporins in having Cys residues and lacking an RGD motif.     

Cellular distribution of okadaic acid in the digestive gland of Mytilus galloprovincialis (Lamarck, 1819)

The distribution of okadaic acid between the digestive and the secretory cells of the digestive gland of Mytilus galloprovincialis was studied for the main purpose of determining if they might be responsible for the two-compartment depuration kinetics found in previous studies. The two cell types did not accumulate okadaic acid to the same degree. However, the concentrations found in each cellular type were not consistent with those expected from the output of the two-compartment depuration model, suggesting that a mechanism other that the differential accumulation in cellular types is involved. Binding to some yet undetermined cellular components is suggested.     

Complete primary structure of statherin,a potent inhibitor of calcium phosphate precipitation,from the saliva of the monkey,Macaca arctoides

Human saliva, which is supersaturated with respect to basic calcium phosphate salts, is stabilized primarily by the presence of two classes of phosphoproteins, statherin and the acidic proline-rich proteins (PRP). These molecules act by inhibiting both primary (spontaneous) precipitation of calcium phosphates in saliva and secondary (surface induced) precipitation of these salts onto dental enamel. The complete amino-acid sequences of several human PRP and the N-terminal sequence of PRP from saliva of M. arctoides have been determined. Similarly, the complete sequence of statherin from human and M. fascicularis saliva is known. We now report the complete structure of statherin from the saliva of the stump-tailed monkey, M. arctoides. The structure was determined by gas-phase sequencing of intact statherin, elucidating positions 1-26, and sequencing an unpurified mixture of tryptic peptides which elucidated the remaining positions through the C-terminus (residue 42) of the molecule. This latter degradation produced an eight amino-acid overlap with that of intact statherin and was confirmed by C-terminal analysis and amino-acid composition of native statherin. The complete amino-acid sequence of M. arctoides statherin is: NH₂-Asp-PSer-PSer-Glu-Glu₅-Lys-Phe-Leu-Arg-Arg₁₀-Leu-Arg-Arg-Phe-Asp₁₅-Glu-Gly-Arg-Tyr-Gly₂₀-Pro-Tyr-Gln-Pro-Phe₂₅-Val-Pro-Pro-Pro₂₉Leu₃₀-Tyr-Pro-Gln-Pro-Tyr₃₅-Gln-Pro-Tyr-Gln-Pro₄₀ -Gln-Tyr-COOH This sequence differs from human statherin at positions 11, 12, 15, 16, 18, 25-27, 38-40 and from M. fascicularis statherin at positions 26 and 28.     

The use of LP533401 as a therapeutic option for renal osteodystrophy affects,renal calcium handling,vitamin D metabolism,and bone health in uremic rats

Background: Klotho is a key regulator of phosphate and Ca²⁺-transport in the kidney. Recently, we showed that treatment with LP533401 improved bone health in rats with chronic kidney disease (CKD) via the normalization of serum phosphate resulting from the reduced renal expression of phosphate cotransporters, including Klotho.

Methods: We evaluated the effect of LP533401 therapy on Klotho-expression-dependent Ca²⁺-transporters, renal calcium handling, and the potential consequences for the bone of uremic rats.

Results: Treatment with LP533401 and its vehicle resulted in the inhibition of transient receptor potential vanilloid receptor subtypes 5 and 6 (TRPV5, TRPV6) and calbindin (CaBP-28k, CaBP-9k) expression. The compensatory acceleration in renal expression of Na+/Ca²⁺-exchanger, 25-hydroxyvitamin d-1α-hydroxylase (CYP27B1), the intensification of vitamin D metabolism, and disruption of sophisticated balance between 1,25-dihydroxyvitamin D–serotonin was observed, especially in rats treated with LP533401. The imbalance between 1,25-dihydroxyvitamin D–serotonin levels led to intensified bone remodeling and improvement in bone geometry, mineral status, and strength in animals treated with LP533401.

Conclusion: The modulation of circulating serotonin and its relation to other regulators of calcium handling can play an important role in calcium homeostasis and bone integrity in CKD rats treated with LP533401.     

Specific expression of an A-kinase anchoring protein subtype,AKAP-150, and specific regulatory mechanism for Na(+),K(+)-ATPase via protein kinase A in the parotid gland among the three major salivary glands of the rat

We have examined the expression of A-kinase anchoring protein (AKAP) in the three major salivary glands, i.e. the parotid gland (PG), submandibular gland (SMG), and sublingual gland (SLG), of the rat to elucidate the functional relevance between saliva secretion and Na(+),K(+)-ATPase regulation by protein kinase A (PKA)-dependent phosphorylation, since an AKAP subtype, AKAP-150, is known to be involved in the regulation of the ATPase in PG. Although AKAP-150 and its mRNA were clearly detected in the PG, they were hardly detectable in either the SMG or SLG. The membrane-bound form of the RII regulatory subunit of PKA, an index for the total amount of AKAP subtypes and therefore of the anchored PKA holoenzyme, was also undetectable in membranes from the SMG and SLG but was found in the PG; though a substantial and comparable amount of Na(+),K(+)-ATPase was present in all of these membrane preparations. Incubation with [gamma-32P]ATP revealed that Na(+),K(+)-ATPase in the PG membranes was quickly phosphorylated upon the addition of cAMP, whereas the ATPases in the membranes from SMG and SLG were not; though they were readily and equally phosphorylated by the exogenously added PKA catalytic subunit. AKAP-150 in the basolateral membranes of PG acinar cells was co-immunoprecipitated with RII by an anti-RII antiserum; and AKAP-150 and Na(+),K(+)-ATPase were immunohistochemically co-localized predominantly on the basolateral membranes, suggesting a possibility that the ATPase might directly interact with the AKAP to form an ATPase/AKAP/PKA complex or associate with the AKAP, such association being mediated via some scaffolding molecule. Expression of AKAP-150 and quick down-regulation of Na(+),K(+)-ATPase by AKAP-anchored PKA in response to cAMP elevation are characteristics specific to PG among the three major salivary glands, suggesting the presence of PG-specific regulatory mechanisms for saliva production/secretion.     

A broad-spectrum organophosphate pesticide O,O-dimethyl O-4-nitrophenyl phosphorothioate (methyl parathion) adversely affects the structure and function of male accessory reproductive organs in the rat

Methyl parathion (MP) is an organophosphate pesticide used in agriculture, but also illegally used to spray homes and businesses to control insects. The present study was designed to investigate adverse effects of MP on accessory reproductive organs. Male Wistar rats aged 13–14 weeks were treated and sacrificed as follows. Experiment 1: 0.0 (water vehicle), 1.75, 3.5 or 7 mg/kg (i.p.) for 5 days and sacrificed on day 14; experiment 2: 0.0, 0.5 or 1 mg/kg (i.p.) for 12 days and sacrificed on day 130; experiment 3: 0.0, 0.5 or 1 mg/kg (i.p.) for 12 days and sacrificed on day 77; experiment 4: 0.0, 0.75 or 1.5 mg/kg (i.p.) for 25 days and sacrificed on day 17 and experiment 5: 0.0 or 3.5 mg/kg (p.o.) for 25 days and sacrificed on day 17, after the last exposure. The accessory reproductive organs were removed, weighed and processed for histopathological analysis. Structural qualitative changes such as epithelial cell morphology and luminal observations were carried out for each organ in all experiments. Epididymis of one side was homogenized and biochemical estimations of acid phosphatase (ACP), cholesterol, total protein, uric acid, and Vitamin C were conducted by calorimetric methods in experiments 4 and 5. In experiment 1 the organ weights did not change; in experiment 2, the epididymal weight increased (P < 0.001); in experiment 3, the weights of ductus deferens decreased at 1 mg/kg and that of seminal vesicle decreased at both dose-levels (P < 0.001). In experiments 4 and 5, weights of epididymis and prostate decreased, whereas in experiment 5, weights of ductus deferens and seminal vesicle increased (P < 0.05–0.001). The sperm density was normal in control, moderately decreased in experiment 1 at 3.5 and 7 mg/kg; in experiment 2 at 1 mg/kg, and in experiment 5 at 3.5 mg/kg, and severely decreased in experiment 3 at 1 mg/kg and in experiment 4 at both dose-levels. The epithelial necrosis and nuclear pyknosis were seen in experiments 1, 3, 4 and 5, whereas nuclear degeneration was seen in experiment 1 and 4 and germ cells in the lumina of epididymis were seen in experiment 4. The nuclear pyknosis in the ductus deferens was seen in all experiments, except at 1.75 mg/kg in experiment 1 and at 0.5 mg/kg in experiment 3. Brush border disruption in the ductus deferens was seen in experiments 1 and 4; sperms were seen in the lumen in experiment 1 at 7 mg/kg, and in experiments 4 and 5. The vacuoles in the epithelium were seen in experiments 1 and 4 and immature germ cells were seen in the lumen in experiment 4. The ACP and Vitamin C levels decreased in experiment 4 at both dose-levels, and in experiment 5 all biochemical parameters tested found decreased (P < 0.01–0.001). The present results indicate that MP affects the structure and function of accessory reproductive organs in the rat.