Does conjugation strategy matter? Cetuximab-conjugated gold nanocages for targeting triple-negative breast cancer cells.

The efficient targeting of cancer cells depends on the success of obtaining the active targeting of overexpressed receptors. A very accurate design of nanoconjugates should be done via the selection of the conjugation strategy to achieve effective targeted nanoconjugates. Here, we present a detailed study of cetuximab-conjugated nonspherical gold nanocages for the active targeting of triple-negative breast cancer cells, including MDA-MB-231 and MDA-MB-468. A few different general strategies were selected for monoclonal antibody conjugation to the nanoparticle surface. By varying the bioconjugation conditions, including antibody orientation or the presence of a polymeric spacer or recombinant protein biolinker, we demonstrate the importance of a rational design of nanoconjugates. A quantitative study of gold content via ICP-AES allowed us to compare the effectiveness of cellular uptake as a function of the conjugation strategy and confirmed the active nature of nanoparticle internalization in cancer cells via epidermal growth factor receptor recognition, corroborating the importance of the rational design of nanomaterials for nanomedicine.

[Metabolomic profiles of exhaled breath condensate of 39 nickel exposed workers]. / Profili metabolomici del condensato espiratorio di 39 lavoratori esposti a nichel.

Nickel compounds have proven lung carcinogenic effects and their processing involve a large amount of population. The aim of this study was to investigate the metabolomic profiles of Exhaled Breath Condensate (EBC) of a group of nickel exposed workers. Nickel in blood, urine and EBC of 39 workers (electroplaters) and 50 controls was measured. The 10 most nickel exposed workers were chosen for the analysis of EBC metabolomic profiles, matched to controls by gender and smoke habits. All the samples were analyzed using the HPLC-MS/MS system (High-Performance Liquid Chromatography/Mass Spectrometry). The profiles of the spectra obtained by the mass spectrometer (Orbitrap) analysis were processed using the MZmine 2.4 software. Nickel concentrations in EBC of the exposed workers were significantly higher compared to controls (1.39 microg/L; 0.039 microg/L, p = 0.017). The observation of the metabolomic profiles pointed out a significantly different response pattern between the exposed and the controls. This result was further studied by a subsequent processing with the XCMS program: an overexpression of 3 hypothetical substances in controls compared to exposed was detected. Although these data must be considered as preliminary, it has been observed that the mass-to-charge ratio of one of these substances may respond to the Phenylacethylglutamine (PAG) one, whose role in the control of cellular cycle is controversial and uncertain. Even if further studies to confirm these results are necessary, the analysis of the metabolomic profiles in the biological matrices is supposed to provide useful information both in the clinical and in the prevention fields.

Renal hilus paraganglioma: a case report and brief review.

Paraganglioma is a rare tumour that originates from any paraganglia. Among extra-adrenal paraganglioma, renal hilus is a rare location. The authors report a case of a 43-year-old female who was admitted for evaluation of a renal mass detected incidentally by ultrasound imaging. Suspecting malignancy, the patient underwent radical nephrectomy. Upon macroscopic examination, the lesion was located into the renal hilus. Histological study revealed a neoplasm constituted of nests of monomorphic cuboidal cells with basophilic granular cytoplasm and round to oval nuclei. Necrosis was absent. The proliferative index (Mib-1) was very low (< 5%). Immunohistochemical examination revealed reactivity for neuron specific enolase (NSE), chromogranin A and synaptophysin. The final diagnosis was renal hilus paraganglioma. The paper shows the difficulty in diagnostic approaches to paraganglioma in this atypical site.

[Myeloid sarcoma in uterine leiomyoma]. / Sarcoma mieloide in leiomioma del miometrio.

In this case report we present a 44-year-old woman with a 2-weekhistory of vaginal bleeding. Gynaecological examination revealed the presence of a polypoid neoformation in the endometrial cavity with a maximum diameter of 4 cm. Histological analysis showed a classic leiomyoma infiltrated by a second monomorphic, highly undifferentiated neoplasia. Immunohistochemical analysis revealed a negative reaction for cytokeratin, CD10, inhibin, CD99, CD20, CD3, TdT and CD34, and positivity for CD45, MPO, CD68 and CD117. A diagnosis of myeloid sarcoma in myometrial leiomyoma was made. The following days the patient showed the onset of an acute myeloid leukaemia M5a. Forty days after diagnosis the patient died for complications related to immunodeficiency caused by therapy. Especially when a common antibody panel reveals negativity for epithelial, mesenchymal and lymphoid markers, this case underlines the importance of considering myeloid sarcoma in differential diagnosis of undifferentiated tumours arising in an extramedullary site in order to avoid errors and permit optimal therapeutic management.

Effects of 12-month GH treatment on bone metabolism and bone mineral density in adults with adult-onset GH deficiency.

Serum bone-Gla protein (BGP), bone alkaline phosphatase (B-AP), and C-terminal cross-linked telopeptide of type I collagen (ICTP) levels were evaluated in 18 adults with acquired GH deficiency (GHD, 14 males and 4 females, age range: 25-59 yr) before, at 3, 6, 9 and 12 months of rec-GH treatment (0.125 IU/kg/week for the first month, followed by 0.25 IU/kg/week for 11 months) and 6 months after the withdrawal of therapy. Total body bone mineral density (BMD, g/cm2) was measured with dual energy X-ray absorptiometry (Hologic QDR 1000/W) before, at 12 months of GH treatment and 6 months after its withdrawal. Before treatment, BGP (mean+/-SE: 5.1+/-0.4 ng/ml), B-AP (59.4+/-6.5 IU/l), ICTP (3.1+/-0.3 ng/ml) levels of patients were similar to in healthy controls (BGP: 5.4+/-0.1 ng/ml; B-AP: 58.2+/-2.0 IU/l; ICTP: 4.1+/-0.3 ng/ml). GH treatment caused a significant increase of BGP, B-AP, ICTP levels, the maximal stimulation of bone resorption, occurring after 3 months of GH treatment, while the maximal effect on bone formation being evident later (at 6th month). A slight decline in BGP, B-AP, T-AP and ICTP levels occurred at 9-12 months of therapy, although the values remained significantly higher than in basal conditions and with respect to healthy controls. Before treatment, mean total body BMD of patients (1.110+/-0.027 g/cm2, range: 0.944-1.350 g/cm2) was not significantly different (z-score: +0.47+/-0.31, NS) from that observed in healthy controls (1.065+/-0.008 g/cm2, range: 1.008-1.121 g/cm2). GH therapy was associated with a significant reduction of mean total body BMD values (6th month: -1.8+/-0.5%, p<0.01; 12th month: -2.1+/-1.0%, p<0.05 vs baseline), particularly evident in the first six months of treatment. Six months after the withdrawal of GH therapy, BGP (5.9+/-0.5 ng/ml), B-AP (57.3+/-7.0 IU/l) and ICTP (3.2+/-0.1 ng/ml) levels returned similar to those recorded before treatment, while total BMD increased (+1.5+/-0.7, p<0.05), remaining however slightly lower than in basal conditions (-0.6+/-1.2, NS). In conclusion, our study shows that: a) acquired GHD in adulthood is associated with both normal bone formation/resorption indexes and normal total body BMD; b) GH therapy causes a significant rise of bone formation/resorption markers (earlier and greater for bone resorption); c) one-year GH therapy is associated with a reduction of total body BMD values, particularly evident in the first 6 months of treatment; d) the effects of GH therapy on bone turnover are transient, being completely reverted six months after the withdrawal of GH therapy; e) the increase of total body BMD (up to baseline values) after GH withdrawal might be explained as consequence of persisting effects of previous GH stimulation on bone remodeling.

Leukotriene D4-induced activation of smooth-muscle cells from human bronchi is partly Ca2+-independent.

Cysteine-containing leukotrienes (cysteinyl-LTs) are potent bronchoconstrictors and play a key role in asthma. We found that histamine and LTD4 markedly constrict strips of human bronchi (HB) with similar efficacy. However, in human airway smooth-muscle (HASM) cells, LTD4, at variance with histamine, elicited only a small, transient change in intracellular calcium ion concentration. HASM cells express both Ca2+-dependent and -independent isoforms of protein kinase C (PKC) (i.e., PKC-alpha and PKC-alpha ). Western blot analysis showed that PKC-alpha is activated by histamine and, to a lesser extent, by LTD4, whereas only LTD4 translocates PKC-alpha. This translocation was specifically inhibited by the LTD4 antagonist pobilukast. Phorbol-dibutyrate ester (PDBu) (a PKC activator) contracted HB strips to the same extent in the presence as in the absence of extra- and intracellular Ca2+. In the absence of Ca2+, LTD4 contracted HB strips to the same extent as did PDBu, suggesting the involvement of a Ca2+-independent PKC in LTD4-mediated signal transduction. PDBu-induced desensitization and the PKC inhibitor H7 abolished the slow and sustained LTD4-triggered contraction of HB strips in the absence of Ca2+, although H7 did not greatly affect the response in the presence of the ion. Thus, in human airways, we identified a novel LTD4 transduction mechanism linked to bronchial smooth-muscle contraction, which is partly independent of Ca2+ and involves the activation of PKC-alpha.

[Solitary cyst of the liver]. / La cisti solitaria del fegato.

The rareness of the illness and the infrequency of infectious complications have led the authors to report a case which has come to their attention. After a brief exam of the etiopathogenesis and pathologic anatomy of these hepatic forms of cyst and the following considerations on their clinical aspects, which range from complete lack of symptoms to the compression of nearby organs (gall ducts, stomach, kidney) due to an increase in their volume, to occurrence of complications (rupture, haemorrhage, suppuration), the authors evaluate the therapeutic problem. Emptying and internal drainage being nowadays discarded and everyone agree on the necessity of excising the whole cystic wall with the minimum loss of hepatic parenchyma, the most indicated operation is total cystectomy. As such an operation is not always feasible because of the adherences of the cyst to the hepatic parenchyma; partial cystectomy can be practiced, leaving in situ part of the cystic wall. If the cyst is complicated by rupture, haemorrhage or suppuration, typical or atypical hepatic resection is recommended. The reported case concerns a patient in good general conditions, bearer of a large left hepatic lobe cyst, complicated by suppuration, with normal liver function, who has been submitted to ablation of the third hepatic segment according to the Ton That Tung transparenchymal technique. Finally the authors point out the possibility of a new conservative therapeutic approach, practiced in a few centres, consisting in echo or CT guided percutaneous needle suction and sclerosis of the cyst with 95% pure alcohol.

Effects on bone mineral density of 12-month goserelin treatment in over 40-year-old women with uterine myomas.

We evaluated the effects on bone mineral density (BMD) of a 12-month treatment with goserelin depot, a gonadotropin-releasing hormone agonist, in a group of women with symptomatic uterine myomas requiring hysterectomy. Sixteen women, mean age 45.6 +/- 5.0, reporting menorrhagia associated with uterine myomas, candidates for hysterectomy, were scheduled to be treated with goserelin depot for 12 months. BMD was measured at the vertebral (L2-L4) and proximal femur level (femoral neck and trochanter) at the start of therapy and 6, 12, and 18 months later using dual energy X-ray absorptiometry (Hologic QDR 1000/W). The patients were followed for a minimum of 6 months after the end of treatment. Thirteen of the 16 women enrolled completed the treatment and three suspended it after 5, 6, and 7 months, respectively, because of side effects (hot flashes, insomnia, depression). Of the 13 women who completed the treatment, three underwent hysterectomy because of myoma regrowth and the recurrence of symptoms 3-18 months later; four reached the menopause 5-16 months later, and six were all menstruating normally with a follow-up varying from 6 to 18 months. After 12 months of therapy we observed a bone loss at vertebral, femoral neck, and trochanter of 4.4% (P < 0.05 versus baseline; P = not significant versus 6 months), 7.5% (P < 0.01 versus baseline, P < 0.01 versus 6 months), and 7.6% (P < 0.001 versus baseline, P < 0.05 versus 6 months), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Age-dependent changes in the mechanisms controlling prolactin secretion and phosphatidylinositol turnover in male rats: effect of phosphatidylserine.

The present study was undertaken in order to better characterize the functional state of anterior pituitary gland in young and old rats by using prolactin secretion and incorporation of radioactive phosphate into phosphatidylinositol (PI) as markers. The in vitro incorporation of radiolabeled phosphate into anterior pituitary PI was significantly (p less than 0.01) greater in young (3-5 months) than in aged (24-25 months) male Sprague-Dawley rats. No significant difference was found in the incorporation by pituitary tissue of 32P into phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Also, the extent of prolactin secretion from isolated pituitary was significantly greater in young than in aged rats, while the prolactin pituitary content was significantly higher in aged animals. In vitro dopamine (DA) decreased the incorporation of 32P into PI, both in young and old pituitary glands, and inhibited prolactin secretion into the incubation medium. Brain cortex-phosphatidylserine (BC-PS), a pharmacologically active purified phospholipid, capable of stimulating the dopaminergic system in the hypothalamus and of decreasing prolactin secretion both in humans and rats in vitro and in vivo, inhibited the incorporation of labeled phosphate into PI of pituitary glands from either young or old rats, but did not alter the prolactin secretion from the glands incubated in vitro. Baseline prolactin plasma levels did not differ significantly between young and old rats either when blood was collected from the trunk after decapitation or underwent sampling from chronically cannulated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Phorbol 12-myristate 13-acetate induces prolactin secretion from rat anterior pituitary gland by the activation of protein kinase-C.

The purpose of this study was to characterize the action of phorbol 12-myristate 13-acetate (PMA), a tumor-promoting agent, on rat anterior pituitary gland, focusing the attention on prolactin secretion. PMA elicited a significant increase in prolactin secretion without affecting phosphatidylinositol turnover, considered as an early post-receptor event controlling PRL secretion. However incubation of anterior pituitary glands with PMA caused a loss of protein kinase-C activity in cytoplasm concomitant with an increased enzyme activity in the membrane. The action of PMA on prolactin secretion seems to be mainly dependent from the redistribution and activation of protein kinase-C. In fact, the phorbol ester did not affect pituitary cAMP and cGMP metabolism either in basal conditions or after theophylline.

[Iatrogenic common bile duct calculosis. Results of choledochorrhaphy with non-absorbable suture material]. / Calcolosi coledocica iatrogena. Esito di coledocorrafia con filo non riassorbibile.

The use of non-absorbable suture material during the surgery of the choledochus can be a cause of choledocholithiasis. An interesting case of a female patient who underwent author's observation for recurrent cholangitis with partial pathological obstruction of the common bile duct after cholecystectomy (1948) and choledochotomy for choledocholithiasis (1977) is reported.

[Non-neoplastic gastro-jejuno-colic fistula. Presentation of 3 cases]. / La fistola gastro-digiuno-colica non neoplastica. Presentazione di 3 casi.

3 cases of non-neoplastic gastrojejunocolic fistula are reported. These were treated by mass resection of the fistulous route with the stomach, small intestine and colon segments involved, followed by immediate restoral of intestinal continuity. After examining the aetiopathogenetic aspects of this disease and reporting the various therapeutic proposals, careful preoperative study is considered vital to clarify the cause of the fistula (hypervagotomy, gastrinoma, hyperparathyroidism). On this basis, surgery is then adapted to avert the danger of recurrence.

[Primary suture of the common bile duct]. / La sutura primaria del coledoco.

The Author reviews 102 cases of primary choledochotomy treated during a 14 years period of biliary surgery; he examines immediate and follow-up results. The excellent results are ascribed parthy for the exactues of indication, parthy to transduodenal sphincteroplasty that had been combined with aforesaid operation in 76% of cases.

[Nuclear insulin assay after a starch and maltose load in chronic diseases of the pancreas: a preliminary note (author's transl)]. / Dosaggio nucleare dell'insulina dopo carico di amido e maltosio nelle pancreatopatie croniche (nota preliminare).

In a group of 48 patients with chronic pancreatitis the authors investigated the variations of blood insulin content after stimulation with starch and maltose, and compared these changes with the correspondind glycemia values. The two curves, practically matching in normal subjects, tend to diverge considerably in patients with chronic pancreatitis. According to the authors, since the assessment of exocrine pancreas function by common laboratory methods is none too satisfactory, this method may yield better appreciation of extant damage to the pancreas.

[Echinococcosis of the pancreas. Description of a case]. / L'echinococco del pancreas. Descrizione di un caso

After a brief survey of the world literature on echinococcus cysts in the pancreas, a case is described. The peculiar clinical picture as well as treatment are underlined.