Surveillance of hospitalised patients with influenza-like illness during pandemic influenza A(H1N1) season in Sicily, April 2009-December 2010.

This paper describes the epidemiology of hospitalised cases with influenza-like illness (ILI) and laboratory-confirmed influenza A cases in Sicily (Italy) during the 2009 influenza pandemic. The first ILI case diagnosed as infected with pandemic influenza A(H1N1)2009 in Sicily was reported in June 2009 and it rapidly became the dominant circulating strain. In the period from 30 April 2009 through 31 December 2010, a total of 2,636 people in Sicily were hospitalised for ILI and 1,193 were laboratory-confirmed for influenza A. Basic demographic and clinical information for all hospitalised patients was collected and population mortality rates (PMRs) and case fatality ratios (CFRs) were calculated. The median age of hospitalised patients infected with pandemic influenza A(H1N1)2009 was significantly lower than that of hospitalised ILI cases in general (18.0 vs. 32.1 years; p<0.0001). Among adults, women were more susceptible than men. The majority of clinical presentations were mild, but 6.6% of hospitalised patients required admission to an intensive care unit, of whom 26.3% had confirmed influenza A. Twenty-four fatal cases were documented. The age group of 45­54 year-olds showed the highest PMRs once hospitalised, while CFRs were higher in elderly patients of 65 years and older. All fatal cases were confirmed as influenza A(H1N1)2009 and most of them had established risk factors for influenza complications.

Cytotoxicity of four types of resins used for removable denture bases: in vitro comparative analysis.

AIM: The aim of this paper was to compare the cytotoxicity of four types of resins used for manufacturing denture bases. METHODS: Nine disk-shaped samples of four resin (two heat-polymerized, one auto-polymerized, and one light-polymerized), 9 samples of glass (negative control) and 9 samples of lead (positive control) were made according to the manufacturer instructions. The materials were tested by contact with BALB/C 3T3 fibroblast cells. Each sample was tested after 24, 48 and 72 hours. The cellular vitality was verified through spectrophotometric analysis of the solution where the colour is directly related to the amount of metabolically active and living cells. The results were analyzed through the one way variance analysis (ANOVA) in order to evaluate significant differences in the behaviour of the resins at 24, 48 and 72 hours. When a significant difference was present, the Games/Howell test for multiple comparisons was used. The significativity level was fixed at P0.05). The light-polymerized resin and the negative control (glass) were so compatible with the cellular carpet that all their values did not show statistically significant differences in any of the three periods of time considered (p>0.05), and their cellular vitality values almost reached the 100%. CONCLUSIONS: The autopolymerized resin showed the major cytotoxicity; the light-polymerized resin, instead, showed an optimal biocompatibility due to the absence of free monomer from its chemical composition.

Cardiac volume overload rapidly induces oxidative stress-mediated myocyte apoptosis and hypertrophy.

Oxidative stress stimulates both growth and apoptosis in cardiac myocytes in vitro. We investigated the role of oxidative stress in the initial phases of cardiac remodeling induced in an animal model by volume overload. As plausible candidates for a connection between oxidative stress and cardiomyocyte apoptosis or hypertrophy, we explored the behaviour of two MAPKs, specifically JNK and ERK. At 48 h of overload, the greatest increase in oxidative stress coincided with a peak of cardiomyocyte apoptosis. This was possibly induced through the mitochondrial metabolism, as evidenced by the release of cytochrome c and a significant increase in the active forms of caspase-9 and -3, but not caspase-8. Oxidative stress markers significantly decreased at 96 h of overload, combined with a marked attenuation of apoptosis and the appearance of hypertrophy. The highest levels of JNK and the lowest levels of ERK phosphorylation were observed at 48 h of overload. Conversely, a sharp increase in ERK phosphorylation was detected at 96 h of overload coinciding with the hypertrophic response. Together these results show that oxidative stress is an early and transient event in myocardial volume overload. They suggest that oxidative stress mediates amplitude dependent apoptotic and hypertrophic responses in cardiomyocytes through the selective activation of, respectively, JNK and ERK.

Possible role of acylphosphatase, Bcl-2 and Fas/Fas-L system in the early changes of cardiac remodeling induced by volume overload.

To identify early adaptive processes of cardiac remodeling (CR) in response to volume overload, we investigated the molecular events that may link intracellular Ca(2+) homeostasis alterations and cardiomyocyte apoptosis. In swine heart subjected to aorto-cava shunt for 6, 12, 24, 48 and 96 h sarcoplasmic reticulum (SR) Ca(2+) pump activity was reduced until 48 h (-30%), but a recovery of control values was found at 96 h. The decrease in SR Ca(2+)-ATPase (SERCA2a) expression at 48 h, was more marked (-60%) and not relieved by a subsequent recovery, while phospholamban (PLB) concentration and phosphorylation were unchanged at all the considered times. Conversely, acylphosphatase activity and expression significantly increased from 48 to 96 h (+40%). Bcl-2 expression increased significantly from 6 to 24 h, but at 48 h, returned to control values. At 48 h, microscopic observations showed that overloaded myocardium underwent substantial damage and apoptotic cell death in concomitance with an enhanced Fas/Fas-L expression. At 96 h, apoptosis appeared attenuated, while Fas/Fas-L expression was still higher than control values and cardiomyocyte hypertrophy became to develop. These data suggest that in our experimental model, acylphosphatase could be involved in the recovery of SERCA2a activity, while cardiomyocyte apoptosis might be triggered by a decline in Bcl-2 expression and a concomitant activation of Fas.

Human herpesvirus type 8 DNA sequences in biological samples of HIV-positive and negative individuals in Sicily.

OBJECTIVE: To evaluate the circulation of a new human herpesvirus (HHV), HHV-8 or Kaposi's sarcoma (KS)-associated herpesvirus in a geographical area where a high incidence rate of classical KS was already present before the appearance of the AIDS epidemic. DESIGN AND METHODS: The study was carried out by analysing: (i) bioptic samples from classic, AIDS-associated KS, and controls; (ii) peripheral blood mononuclear cells (PBMC) from classic KS, HIV-positive subjects with and without KS and healthy HIV-negative individuals; (iii) semen samples from heterosexual HIV-positive and HIV-negative individuals affected or not by KS; and (iv) cervical swabs from HIV-negative healthy heterosexual females. All specimens were tested for the presence of HHV-8 DNA sequences by a two-step polymerase chain reaction. RESULTS: Positive results were obtained in 90% of bioptic samples of classic KS and in 100% of AIDS-associated KS. Viral sequences were also present in 50% of PBMC of subjects with classic KS and AIDS-associated KS, in 10% of AIDS patients without the angiosarcoma and in 11% of healthy HIV-negative individuals. Finally, HHV-8 DNA was detected in 13% of semen of HIV-negative heterosexual individuals and in 10% of AIDS patients without KS. Both PBMC and ejaculates from the same individual gave positive results. No HHV-8 sequences were found in cervical swabs. CONCLUSIONS: HHV-8 is widespread in the general population in Sicily since it was detected in PBMC and semen of heterosexual HIV-negative individuals and is not found only in high-risk groups. The viral load appears to be more elevated in a high-risk population and it may be ascribed to a viral reactivation. The higher incidence rates of KS in Sicily compared with northern Italy and other European countries might be related to the presence of HHV-8 in the general population.

Antibodies to human herpes virus type 8 (HHV8) in general population and in individuals at risk for sexually transmitted diseases in Western Sicily.

BACKGROUND: Human herpes virus 8 (HHV8) appears to be the primary aetiologic agent of Kaposi sarcoma (KS). To study the distribution of HHV8, a seroepidemiological study was carried out in western Sicily, where a high incidence rate of classical KS is well documented. METHODS: A total of 970 sera of healthy human immunodeficiency virus (HIV) negative individuals of general population (1-70 years old) and 742 sera of individuals in different risk groups for HIV infection were evaluated by means of an indirect immunofluorescence assay able to detect antibodies to lytic and latent HHV8 antigens. RESULTS: Crude seroprevalence to HHV8 antigens was 11.5% in the general population, and it increased significantly with age from 6% under age 16 to 22% after age 50. Significantly higher HHV8 seroprevalence rates were detected among HIV positive and negative homosexual men (62% and 22%, respectively), men who had sex with prostitutes (40% and 29%, respectively); female prostitutes (42% and 30%, respectively), and clients at a sexually transmitted disease clinic (male: 60% and 33%, respectively, female: 63% and 43%, respectively). In contrast, heterosexual intravenous drug users had seroprevalence rates comparable to those found in the general population. CONCLUSIONS: The results suggest that HHV8 infection is widespread in Western Sicily. The high seroprevalence in individuals with high risk sexual activity point to the role of sexual behaviour in the transmission of the infection in adults, whereas the detection of antibodies in younger population (under 16 years old) is suggestive of a non-sexual route of transmission, probably occurring during childhood by close personal contact.

Altered Cx43 expression during myocardial adaptation to acute and chronic volume overloading.

Gap-junctions are specialized regions of intercellular contacts allowing electrical impulse propagation among adjacent cardiomyocytes. Connexin43 (Cx43) is the predominant gap-junction protein in the working ventricular myocardium and its reduced expression has been extensively implicated in the genesis of conduction abnormalities and re-entry arrhythmia of chronically hypertrophied hearts. In contrast, data on the role played by this protein during cardiac remodeling and early phases of developing hypertrophy are lacking. Therefore, in the present study, we investigated this issue using an experimental model of pig left ventricle (LV) volume overloading consisting in the creation of an aorto-cava fistula. At scheduled times (6, 24, 48, 96, 168 h, and 2, 3 months after surgery) echocardiographic and haemodynamic measurements were performed and myocardial biopsies were taken for the morphological and biochemical analyses. When faced with the increased load, pig myocardium underwent an initial period (from 6 up to 48 h) of remarkable tissue remodeling consisting in the occurrence of cardiomyocyte damage and apoptosis. After that time, the tissue developed a hypertrophic response that was associated with early dynamic changes (up-regulation) in Cx43 protein expression, as demonstrated by Western blot and confocal immunofluorescence analyses. However, an initial transient increase of this protein was also found after 6 h from surgery. With the progression of LV hypertrophy (from 168 hr up to 3 months), a reduction in the myocardial Cx43 expression was, instead, observed. The increased expression of Cx43 protein during acute hypertrophic response was associated with a corresponding increase in the levels of its specific mRNA, as detected by RT-PCR. We concluded that up-regulation of Cx43 gap-junction protein could represent an immediate compensatory response to support the new working conditions in the early stages of ventricular overloading.

Intermittent reperfusion extends myocardial preservation for transplantation.

A protocol was developed to compare prolonged heart preservation by hypothermic storage with prolonged hypothermic storage interrupted by a period of reperfusion. Hearts from adult mongrel dogs were excised after administration of 4 degrees C crystalloid cardioplegia. Group A hearts (N = 7) underwent 7.5 hours of ischemia at 4 degrees C followed by 1.5 hours of reperfusion and rewarming (A0). Group B hearts (N = 8) underwent 3 hours of ischemia at 4 degrees C, 1.5 hours of reperfusion and rewarming (B1), 3 additional hours of ischemia at 4 degrees C following repeat cardioplegia, and finally 1.5 hours of reperfusion and rewarming (B2). During reperfusion, hearts were defibrillated and left ventricular (LV) function was assessed by measuring isovolumic peak systolic pressure and maximum positive rate of rise of LV pressure (+dP/dtmax) with an intraventricular balloon. LV biopsy samples for adenosine triphosphate (ATP) assay were obtained before ischemia and after each rewarming period. One Group A heart could not be defibrillated and studied. All Group B hearts completed the protocol. LV function, as assessed by peak pressure and +dP/dtmax, at B1 and B2 exceeded values obtained at A0, but the differences were not statistically significant. The mean ATP level was 63.4 +/- 7.7% of baseline at B1 and 79.7 +/- 4.3% of baseline at B2 (p less than .03). The mean ATP level was 57.9 +/- 5.9% of baseline at A0 (p less than .007, B2 vs. A0). It is presumed that intermittent reperfusion allows repletion of substrate stores, which results in improved myocardial protection.(ABSTRACT TRUNCATED AT 250 WORDS)

High-dose aprotinin: hemostatic effects in open heart operations.

Two groups of patients were prospectively studied to evaluate the hemostatic effects of high-dose aprotinin in open heart operations. Group A patients (n = 22) received aprotinin during the entire surgical procedure. Group B patients (n = 12) served as controls. The groups were homogeneous for base variables and for cardiopulmonary bypass duration. Postoperative bleeding was lower in group A (mean, 486 mL) than in group B (830 mL) (p less than 0.01). The need for banked blood decreased by approximately half in the aprotinin patients (mean: group A, 213 mL; group B, 409 mL). Hemoglobin levels were similar in the two groups (postoperative day 7: group A, 11.29 g/100 mL; group B, 11.26 g/100 mL; NS). Platelet count decreased at the end of operation in both groups (99,000 and 95,000/mL, respectively; NS) and then increased beyond baseline levels before discharge. No complications were observed that could be attributed to aprotinin. In conclusion, we believe that the use of high-dose aprotinin is safe and effective. It decreases blood loss and reduces the need for banked blood in cardiac operations, particularly in select groups of patients (eg, those undergoing reoperation, Jehovah's Witnesses, those with renal failure).

Are macrophages involved in early myocardial reperfusion injury?

BACKGROUND: Neutrophils are the predominant phagocytes in the early stages of myocardial ischemia-reperfusion response and are also implicated in the development of tissue damage. This study examined the role of recruited macrophages in the evolution of this tissue injury. METHODS: Farm pigs were subjected to 30 minutes of myocardial ischemia followed by 30 minutes of reperfusion. Biopsy samples were taken from the control, ischemic, and ischemic-reperfused left ventricle wall and processed for both morphologic and biochemical analyses. In situ production of tumor necrosis factor-alpha was evaluated by Western blot and immunofluorescence. A full hemodynamic evaluation was also performed. RESULTS: Myocardial ischemia and early reperfusion caused marked neutrophil and macrophage tissue accumulation and tumor necrosis factor-alpha production by the injured tissue. Immunofluorescence studies allowed us to localize tumor necrosis factor-alpha predominantly in tissue-infiltrating macrophages. No depression in the global myocardial contractile function was observed, either during ischemia or after reperfusion. CONCLUSIONS: These data suggest that the newly recruited macrophages within the ischemic and early post-ischemic myocardium may play a role in promoting neutrophil tissue infiltration and subsequent neutrophil-induced tissue dysfunction by producing tumor necrosis factor-alpha.

Clinically relevant cognitive impairment after cardiac surgery: a 6-month follow-up study.

BACKGROUND AND PURPOSE: The majority of studies on neuropsychological complications after cardiac surgery used the raw variation of selective tests scores to define the occurrence of cognitive decline. We prospectively estimated the frequency of cognitive impairment after cardiac surgery, with a particular emphasis on persistent and clinically relevant cognitive decline. Possible baseline and operative predictors were also evaluated. METHODS: An extensive neuropsychological battery was administered to 110 patients (mean age 64.1+/-9.4 years; 70.9% males) undergoing cardiac surgery before and 6 months after the operation. After evaluating the variations in the cognitive performances, two independent neuropsychologists ranked the patients as unchanged-improved, mildly-moderately deteriorated, or severely deteriorated, using a global and functionally oriented judgement. The degree of the impairment was determined in relation to its impact on everyday life activities. RESULTS: Ten patients (9.1%) were ranked as severely deteriorated, 22 (20%) as mildly-moderately deteriorated, and 78 (70.9%) as unchanged-improved. Cognitively impaired patients were older (p=0.031), more often females (p=0.005), with a low education level (p=0.013). At multivariate analysis, female gender (odds ratio (OR) 6.14, 95% confidence interval (95% CI) 2.16-17.50), baseline use of beta-blockers (OR 4.55, 95% CI 1.30-15.92), and PaO2 at arrival in intensive care unit (OR for 1 mm Hg increment 1.012, 95% CI 1.004-1.020) were significant predictors of cognitive impairment of any degree. Positive predictors of severe cognitive impairment were history of hypertension (OR 5.33, 95% CI 1.03-27.64) and PaO2 at arrival intensive care unit (OR for 1 mm Hg increment 1.020, 95% CI 1.006-1.035), while education was protective (OR per year of increment 0.53, 95% CI 0.31-0.90). CONCLUSIONS: A considerable proportion of cardiac surgery patients may undergo clinically relevant cognitive impairment. The knowledge of variables influencing cognitive outcome is essential for the adoption of preventive measures.

Cardiac tamponade secondary to intrapericardial rupture of a hepatic amoebic abscess.

We report the case of a patient who underwent an emergency subxiphoid pericardiectomy with evacuation of 1700 cc of anchovy sauce-like fluid. On pericardial inspection a fistulous communication was seen with a cavity in the left hepatic lobe. No loculi or adhesions were found in the pericardial sac and no further surgical procedure was advised. The hepatic cavity and pericardium were drained: bacteriological investigation of the fluid yielded entamoeba histolytica and chloroquine was administered. Apart from the rarity of the observation in western countries, we discuss surgical approaches in cases of unknown purulent pericardial effusions, stressing the possibility of achieving a good surgical result even with a minor surgical procedure such as subxiphoid pericardiectomy and drainage.

Intramural hematoma of the aorta: diagnosis and treatment.

OBJECTIVE: Increasing use of modern high-resolution imaging techniques yields to describe very early stages of aortic pathology which, if left untreated, may lead to overt aortic dissection. One typical example is aortic intramural hematoma (IMH) with a limited number of cases described in the literature and uncertainties still existing about the most appropriate treatment. Purpose of our study is to report our experience in the evaluation and treatment of IMHs. METHODS: From 1991 to 1999 175 patients were conveyed to our centre for aortic dissection; in nine of them diagnosis of acute IMH was performed. RESULTS: Diagnosis was obtained by means of conventional CT scan of the chest. All the patients underwent surgery, one patient died (11%). At the follow-up (mean 31 months) eight patients were alive and well and did not require any other cardiac surgery. CONCLUSIONS: The possibility to progress to overt aortic dissection may explain the need to an early diagnosis in the treatment of acute IMHs. Immediate surgical treatment is, in our experience, the preferred therapeutic option.

Partial anomalous pulmonary venous return of an uncommon type.

We report a rare case of complete anomalous venous return from the right lung to the supradiaphragmatic part of the inferior vena cava, associated with sinus venosus atrial septal defect. The defect was corrected with direct reimplantation of the anomalous venous trunk into the left atrium and direct suture of the ASD. A good operative result was obtained. As a great variety of associative patterns between partial APVR and ASD exists in literature, we outline the necessity of an accurate pre- and intra-operative diagnosis, in order to achieve the most satisfactory surgical result.

Repair of coarctation of the thoracic aorta without resection. Patch graft aortoplasty. Follow-up study of 46 cases.

Two hundred and seventy-six patients (276) over the age of one were operated for coarctation of the thoracic aorta from 1949 to 1972. Patch graft aortoplasty, rather than resection of the coarcted segment was the operation of choice in forty-six patients. No hospital or late deaths occurred in this group of patients. Associated defects were present in 20% of these cases. The first patch aortoplasty was performed in 1962. The follow-up study covers a period of 12 years. Haemodynamic study and aortography were performed to assess the long term results. There was no evidence of recoarctation in the follow-up period up to 14 years following surgery and the pressure difference across the repair varied between a minimum of 5 mmHg to 18 mmHg (Table 2). The technique of patch aortoplasty without resection as been extended to a large number of patients and seems to be a very important factor in avoiding a circular constricting anastomosis, preventing the growth of the anastomosis. Maintaining the whole wall of the aorta may also influence the further growth and prevent recurrence in infants and children. Very little surgical dissection is required and suture line tension is also avoided. The technique of patch aortoplasty should be recommended in infants, children and adults with difficult and hazardous dissection.

Inhibition of human immunodeficiency virus 1 (HIV-1) by variant B of human herpesvirus 6 (HHV-6).

Four HHV-6 strains were initially isolated during attempts to observe HIV-1 replication in cultured primary lymphocytes from 48 patients with AIDS. HHV-6 DNA from each strain was extracted from primary cell cultures and amplified using specific primers in a nested polymerase chain reaction (PCR) assay. All HHV-6 strains were classified as B variants by submitting the PCR products to the digestion of two restriction enzymes (Hind III and Bgl II). Since in primary cultures, the appearance of HHV-6 cytopathic effect was followed by a progressive reduction of HIV-1 replication, we tried to reproduce the observed inhibition in vitro. Two HHV-6 strains, used throughout the experiments, showed their ability to suppress HIV-1 replication when the viruses co-infected CD4+T lymphocyte cultures. While the intrinsic mechanism of this finding still remains unclear, the inhibition of HIV-1 replication was observed only when a high multiplicity of infection (m.o.i.) of HHV-6 and a low m.o.i. of HIV-1 were used in dually infected cell cultures. By using a semiquantitative determination of HIV-1 cDNA by PCR, it appears that the inhibition begins in infected cell cultures and, once established, does not allow any further HIV-1 replication.

Distribution of hepatitis C virus genotypes among intravenous drug users. A ten-year study in Palermo, Sicily.

A cross-sectional study was carried out on the sera of 88 active intravenous drug users (IVDU) collected between 1985 and 1986 to evaluate the circulation of HCV genotypes in Western Sicily. The patients were grouped by age and classified by their HIV status. Genotype 3a (48.8% of all cases) was most frequently detected, followed by genotype 1a (20.4%) and type 1b (17.0%). No significant differences in HCV genotype distribution were observed between HIV positive and negative individuals. Next, the HCV genotype distribution found in sera samples of IDVUs drawn between 1985 and 1986 was studied and divided into three age groups. The genotype distribution in the younger group was then compared with samples collected ten years later, between 1995 and 1996, from young HIV negative IVDU individuals. A different distribution between HCV genotypes 3a and 1a was found with a relative, though not significant, increase in the detection of genotype 1a (38%). Finally, sera from six IVDUs obtained at three different times over a ten-year period were genotyped for HCV. None of the subjects showed any change in the genotype found at the first sampling throughout the ten years. The results suggest that a) genotype 1a and 3a are the most common among IVDUs in Western Sicily, b) concurrent HIV infection does not seem to influence HCV genotype and c) infected IVDUs harbor almost exclusively one genotype.

Ultrastructural evidence of myocardial alterations in the course of heterotopic heart transplantation.

Using a novel model of heterotopic rat heart transplantation, the present study was undertaken to evaluate whether parenchymal and microvascular alterations of the ischemic and reperfused myocardium occurred and could be related to local neutrophil infiltration. In such a model, hearts were rapidly excised from donor rats, maintained in a cold saline solution at 4 degrees C and then reimplanted in recipient animals. Muscle biopsies of the ischemic and reperfused myocardium were analysed by ultrastructural and immunohistochemical techniques. Although the cold storage of the hearts provided a good protection against the ischemic insults, reperfusion with the recipient blood caused severe myocardial cell injury and microvascular damage. In particular, the microvascular endothelium showed numerous discontinuities due to the partial destruction of endothelial cell. The altered endothelial integrity was associated with aggregation and adhesion of platelets to the luminal surface. Contrary to other models of ischemia and reperfusion, where neutrophils are considered the major source of oxygen radicals and cellular dysfunctions at reperfusion, in our samples the burst of these toxic metabolites did not originate from such cells. In fact, no neutrophils were seen to accumulate within the ischemic as well as reperfused myocardium. Accordingly, the microvascular endothelium did not express E-selectin, an adhesive molecule which is responsible for the increased adherence and emigration of neutrophils in the microvasculature.

Dynamics and molecular evolution of HIV-1 strains in Sicily among antiretroviral naïve patients.

HIV-1 subtype B is the most frequent strain in Sicily. To date, there is no available data about the genetic diversity of HIV-1 viral strains circulating in Sicily among antiretroviral (ARV) naïve subjects and the role of immigration as potential determinant of evolutionary dynamics of HIV-1 molecular epidemiology. For this purpose, HIV-1 polymerase (pol) sequences obtained from 155 ARV naïve individuals from 2004 to 2009 were phylogenetically analysed. The overall rate of HIV-1 non-B infections was 31.0% (n=48/155), increasing from 7.8% in 2004-2006 to 40.9% in 2009, and about one-third were identified as unique recombinant forms. CRF02_AG was the prevalent non-B clade (n=28/48, 58.3%), while subtype C-related strains were responsible for about 30% HIV-1 infections. Non-B viruses strictly associated with heterosexual transmission (85.4%) and were mostly found among immigrants (77.1%). Phylogenetic analysis of non-B sequences found in foreign-born subjects was geographically correlated to the respective country of origin. Moreover, the detection of non-B viral variants in the autochthonous population may support an increasing genetic diversity in Sicily as well as a local circulation of HIV strains also uncommon in our country. In Sicily, HIV-1 epidemic is still mostly attributable to the B subtype. Nevertheless, migration and population movements are progressively introducing novel HIV-1 subtypes causing a continuous increase of HIV-1 molecular dynamic at local level. Molecular surveillance is needed to monitor the genetic evolution of HIV-1 epidemic.