This study reports on a collection of spiders from Misallatah Nature Reserve and National Park (MNRNP), north western Libya, including five species recorded for the first time in the country. 75 spider samples (13 adult males, 20 adult females, 42 juveniles), comprising 12 families (Agelenidae; Araneidae; Dysderidae; Gnaphosidae; Lycosidae; Palpimanidae; Pisauridae; Philodromidae; Pholcidae; Salticidae; Thomisidae; Zodariidae), 13 genera (Lycosoides Lucas, 1846; Argiope Audouin, 1826; Dysdera Latreille, 1804; Nomisia Dalmas, 1921; Hogna Simon, 1885; Palpimanus Dufour, 1820; Holocnemus Simon, 1873; Aelurillus Simon, 1885; Menemerus Simon, 1868; Plexippus C. L. Koch, 1846; Phlegra Simon, 1876; Bassaniodes Pocock, 1903; Zodarion Walckenaer, 1826) and 13 species (Argiope lobata Pallas, 1772; Dysdera crocata C. L. Koch, 1838; Nomisia recepta Pavesi, 1880; Palpimanus gibbulus Dufour, 1820; Holocnemus pluchei Scopoli, 1763; Plexippus paykulli Audouin, 1826; Bassaniodes lalandei Audouin, 1826; Zodarion pileolonotatum Denis, 1935; Lycosoides flavomaculata Lucas, 1846; Hogna effera O. Pickard-Cambridge, 1872; Aelurillus blandus Simon, 1871; Menemerus taeniatus L. Koch, 1867; Phlegra yaelae Prószyński, 1998) were collected; the last five species are first records in Libya.

Purpose: The first aim of this study was to analyse the presence of circulating Claudin 4 (CLDN4) mRNA in serum and to evaluate whether the rate of CLDN4 mRNA can be used as a tumor marker to discriminate breast cancer patients and healthy women. The second aim was to compare CLDN4 mRNA measured using real-time quantitative RT-PCR with CA 15-3 measured using immunoassay as biomarker for the diagnosis of breast cancer.

Experimental Design: The present study collected serum samples

from 118 patients with recently diagnosed breast cancer and 30 matched healthy women. The number of copies of CLDN4 mRNA and the CA-15-3 rate were analysed and the percentage of patients with increased marker level were compared.

Results: The quality and the stability of mRNA which were evaluated

using the housekeeping RPLPO gene in all studied serum samples were confirmed (p>0.05). The expression level of CLDN4 mRNA was found significantly increased in serum of 64% patients with breast cancer from the threshold calculated in healthy women (p<0.001) versus 6% for CA 15-3. The expression of claudin4 was not correlated with histological type, tumour size, nodal status, tumor grade, and status of ER, PR and Her2 but only with age.

Conclusions:

Our results showed that the expression of CLDN4 mRNA by real-time quantitative PCR give the opportunity to predict breast cancer before pathological diagnosis in a large majority of patients.