Choline kinase inhibition in rheumatoid arthritis.

OBJECTIVES: Little is known about targeting the metabolome in non-cancer conditions. Choline kinase (ChoKα), an essential enzyme for phosphatidylcholine biosynthesis, is required for cell proliferation and has been implicated in cancer invasiveness. Aggressive behaviour of fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) led us to evaluate whether this metabolic pathway could play a role in RA FLS function and joint damage. METHODS: Choline metabolic profile of FLS cells was determined by (1)H magnetic resonance spectroscopy ((1)HMRS) under conditions of ChoKα inhibition. FLS function was evaluated using the ChoKα inhibitor MN58b (IC50=4.2 µM). For arthritis experiments, mice were injected with K/BxN sera. MN58b (3 mg/kg) was injected daily intraperitoneal beginning on day 0 or day 4 after serum administration. RESULTS: The enzyme is expressed in synovial tissue and in cultured RA FLS. Tumour necrosis factor (TNF) and platelet-derived growth factor (PDGF) stimulation increased ChoKα expression and levels of phosphocholine in FLS measured by Western Blot (WB) and metabolomic studies of choline-containing compounds in cultured RA FLS extracts respectively, suggesting activation of this pathway in RA synovial environment. A ChoKα inhibitor also suppressed the behaviour of cultured FLS, including cell migration and resistance to apoptosis, which might contribute to cartilage destruction in RA. In a passive K/BxN arthritis model, pharmacologic ChoKα inhibition significantly decreased arthritis in pretreatment protocols as well as in established disease. CONCLUSIONS: These data suggest that ChoKα inhibition could be an effective strategy in inflammatory arthritis. It also suggests that targeting the metabolome can be a new treatment strategy in non-cancer conditions.

Determination of water content by capillary gas chromatography coupled with thermal conductivity detection.

This article presents some experience obtained by applying capillary gas chromatography coupled with thermal conductivity detection (GC/TCD) to the determination of water in substances for pharmaceutical use. This technique represents a useful, orthogonal tool complementary to water determination methods based on volumetric or coulometric titration. It can also represent an alternative technique when such titrations are not applicable. This article presents the preliminary results obtained in a number of case studies where a GC/TCD procedure was applied in comparison with pharmacopoeial methods to substances with different water contents.

Cadmium biosorption on Spirulina platensis biomass.

Dry biomass of Spirulina platensis re-hydrated for 48 h was employed as a biosorbent in tests of cadmium(II) removal from water. Various concentrations of biomass (from 1 to 4 g l(-1)) and metal (from 100 to 800 mg l(-1)) were tested. Low biomass levels (X(o)2 g l(-1)) ensured metal removal up to 98% only at Cd(0)= 100 and 200 mg l(-1), while X(o)2.0 g l(-1) were needed at Cd(0)=400 mg l(-1) to achieve satisfactory results. Whereas X(o)=4.0 g l(-1) was effective to remove up to Cd(0)=500 mg l(-1), a further increase in metal concentration (Cd(0)=600 and 800 mg l(-1)) led to progressive worsening of the system performance. At a given biomass levels, the kinetics of the process was better at low Cd(2+) concentrations, while, raising the adsorbent level from 1.0 to 2.0 g l(-1) and then to 4.0 g l(-1), the rate constant of biosorption increased by about one order of magnitude in both cases and the adsorption capacity of the system progressively decreased from 357 to 149 mg g(-1).

Inter-laboratory reproducibility of the LC-ELSD method for the control of composition of sesame oil recently introduced in the Ph. Eur.

Evaporative light-scattering detection (ELSD) is an increasingly important technology for the control of substances for pharmaceutical use. A LC-ELSD method has recently been introduced in the Ph. Eur. monograph for sesame oil, refined to control the composition of triglycerides. A collaborative study was run to assess the reproducibility of the new method and to confirm that it was suitable for pharmacopoeial use.

Relationship between intrathyroid calcifications and thyroglobulin in endemic goiter.

Intrathyroid calcifications represent a common finding within simple or nodular goiters, but, as far as they can be found also inside papillary and medullary thyroid carcinomas, an ultrasonographic detection of intrathyroid calcifications stands as a different diagnosis problem. We have been looking for the presence of parameters associated with thyroid calcifications in patients affected by simple or nodular goiter, either sporadic or endemic. We studied 284 euthyroid subjects, 250 females, ageing from 24 to 90 years, affected by a simple goiter, in the 9.51% of the cases, and by a nodular goiter in the remaining part. 69.37% of the patients came from an endemic goiter area, while the others were affected by sporadic goiter. We tested fT3, fT4, TSH, hTG, Ab-TG, Ab-TPO and performed an ultrasonography in all the subjects, 57.75% of patients shown intrathyroid calcifications in the 57.75% of them. We applied a multistep discriminant analysis taking for the presence/absence of calcifications as dependent variable and we tried to find which variable, by itself or in combination with others, could foretell its presence. We also created a new variable (TG1) to differentiate normal from supraphysiologic concentrations of hTG (< 60 ng/ml). The variable with the highest significance F originated from endemic goiter area (F = 96.36), followed by TG1 (F = 24.46) and age (F = 10.61). On the contrary hTG did not relate to calcifications, due to non-proportionally direct relationship between these two parameters, afterwards we used the multistep logistic regression that gave overlapping significances. This means that supraphysiologic hTG rates are sufficient to predict the possible presence of intrathyroid calcifications. In conclusion, as far as a follicular hyperstimulation can be assumed, especially if long-lasting, the presence of intrathyroid calcifications should rise a clinical suspect toward an old goiter rather than a neoplastic lesion.

In vitro stability of polymerase chain reaction-generated DNA fragments in serum and cell extracts.

The potential use of polymerase chain reaction (PCR)-generated DNA fragments (PCR-DNAs) as pharmaceutical agents has previously been suggested, with the demonstration of the in vitro cellular internalization and biologic activity of PCR-DNA decoy molecules targeted to human estrogen receptor gene. In order to provide information on the stability of these double-stranded DNA molecules, the nuclease resistance of PCR-DNAs of different sizes was studied in different conditions and experiments. Simulating in vitro and in vivo transfection protocol, we demonstrated that PCR-DNAs exhibited good stability toward fetal bovine serum (FBS) and adult human serum nuclease digestion. In addition, when the protective activity of liposome-based formulations toward nuclease digestion was tested, it was shown that the stability of PCR-DNAs could be further increased (up to 7 days) when a liposome-mediated delivery system was employed.

An immunohistochemical study of progesterone receptor in thyroidal Hürthle cells tumors.

Estrogen (ER) and progesterone (PgR) receptors are commonly represented on normal and neoplastic thyroid tissues, while a few data are available about their expression in Hürthle cells tumors. Seventeen patients who underwent thyroidectomy for a neoplasia that showed oxyphilic elements were studied by an immunohistochemical method and the presence of PgR in the 30% of them, was demonstrated, while no ER expression could be found. We tried, therefore, to establish a correlation between these proteins and age, sex or indexes of the supposed estro-progestinic concentration of our patients. None of these factors resulted significantly related to the receptors expression. Since PgR are expressed in the oxyphilic cells neoplasms, further studies on both ER, and PgR are needed in order to point out their possible biological importance.

Epidemiological, clinical and allergological observations on pompholyx.

We have studied a group of 104 patients with pompholyx, to investigate the relationship between allergological factors and its etiopathogenesis. The following examinations were performed: blood sampling (routine tests and IgE levels), allergological tests (patch, prick, intradermal, and oral provovation tests with nickel sulphate), skin biopsy to exclude pemphigus vulgaris or bullous pemphigoid. An accurate history of familial and personal allergic diathesis was enquired for and various possible aggravating factors (season, microclimate, perspiration and emotional stress) were considered. The results were age and sex-matched with a healthy control group (208 subjects). We found familial and personal atopic diathesis in 50% of patients versus 11.5% of controls (p less than 0.001); 39 patients (37.49%) also had high levels of IgE. Nickel sulphate was the allergen with the highest positivity on patch testing: 20.19% versus 6.25% of the control group (p less than 0.001). The % of patients allergic to nickel reached 26%, including those (6 patients) reacting to the oral provocation test. Season (43 patients) and hyperhidrosis (38) were the aggravating factors most commonly claimed. We detected no correlation between age, sex, grading of pompholyx and the allergological parameters investigated. Though several different allergological findings have previously been reported in dyshidrosis, their role in its pathogenesis has not yet been fully explained. We think that different haptens or antigens can produce the same clinical and histological picture of pompholyx in predisposed subjects.

Allergy to components of total hip arthroplasty before and after surgery.

Sensitivity to a series of haptens was studied in two groups of patients by means of patch tests. The first group was made up of 66 patients with total hip replacements, 12 of whom had aseptic loosening of the implant. The second group, which was used as a control, consisted of 41 patients scheduled to undergo total hip replacement. Ten patients (15.1%) in the first group tested positive; one of these had aseptic loosening. In the control group, two patients (4.8%) tested positive. The theory that an allergy may be the cause of loosening has a biologic foundation, but it can only be used to explain a few cases. However, since considerable vanadium sensitivity was found, we suggest that this material be used as little as possible in the manufacturing of prosthetic components.

Effect of UVB plus tar therapy on serum levels of interleukin-2 receptors in patients with psoriasis.

Baseline serum levels of interleukin-2 receptor (IL2R) were measured in 65 patients with active psoriasis. IL2R levels in psoriatic patients were significantly higher than in healthy controls (582.4 +/- 289.23 u/ml vs. 369.64 +/- 111.10 u/ml; P less than 0.05), but did not differ statistically from values found in an atopic dermatitis control group (619.88 +/- 254.27 u/ml). Sex, age and severity of the disease do not affect levels of IL2R. The same IL2R levels were measured in 26 psoriatic patients receiving UVB plus tar therapy. This therapy, continued until clinical remission, lowered IL2R levels to values comparable to controls. This decrease may be due to an immunosuppressive effect of therapy.

Incidence of polycystic ovaries in patients with late-onset or persistent acne: hormonal reports.

Forty-six women affected by late-onset or persistent acne were studied in order to investigate the frequency of hormonal abnormalities and polycystic ovaries. Hirsutism, perioral distribution of acne lesions and irregular menses were recorded. Hormonal measurements and ovarian echographies were performed. Twenty-four patients were affected with polycystic ovaries, detected by ultrasound scanning. Among the acne patients, the women with ovarian abnormalities had higher values of androstenedione, dehydroepiandrosterone, dehydroepiandrosterone sulfate and luteinizing hormone (LH), and a higher LHT/follicle-stimulating hormone ratio than those with acne and without ovarian abnormalities. This study indicates the prevalence of polycystic ovaries in women with late-onset or persistent acne. Moreover, hormonal abnormalities indicate a subgroup of acne patients defined by the presence of ovarian disorders.

A comparative study on peripheral blood lymphocyte subpopulations in different kinds of warts.

Peripheral blood T-cell subpopulations were evaluated in 36 patients with clinically different types of warts, subdivided in 4 groups (common, genital, flat and plantar warts). A significant decrease was found in OKT3 and OKT4 subsets total count and in OKT4/OKT8 ratio in patients with common and genital warts as compared with controls. Only in common and genital warts did we also observe a significant decrease of percentage of OKT4 subset. No significant difference of considered parameters was observed in flat and plantar warts as compared to controls, apart from a significant increase in number of OKT8 subset in flat warts. We then discuss this different status of C.M.I. in patients with different clinical warts, stressing the importance of various types of HPV.

[Psoriasis and day hospital. Clinical and therapeutic considerations concerning 104 cases]. / Psoriasi e Day Hospital. Considerazioni cliniche e terapeutiche su 104 casi.

One hundred and four patients with psoriasis, treated in regimen of Day Hospital, have been examined. The diagnosis was widespread plaque type psoriasis (67.3%), guttate psoriasis (18.3%) and nail psoriasis (1.0%). Furthermore psoriatic erythroderma has been observed in 2.9% and pustular psoriasis of Barber in 1.9%. 10% of the patients showed psoriatic arthritis. 71.2% of the patients have been successfully treated with UVB and PUVA irradiation therapy (respectively 86.6% and 70.0% of complete clearing or marked improvement). Etretinate and topical therapy were successful (respectively 67.0% and 70.0% of complete clearing or marked improvement). Day Hospital has proved to be the ideal regimen for the treatment and control of psoriatic patients.

[Sexually transmitted infections in patients with condylomata acuminata]. / Infezioni a trasmissione sessuale in pazienti affetti da condilomi acuminati.

42 patients (26 males, 16 females) with genital and perianal warts were submitted to serological testing for HBV markers, anti HSV antibodies and syphilis. Specimens were also collected for microscopic and cultural examination for Neisseria gonorrhoeae, Chlamydia Trachomatis, Trichomonas vaginalis, Candida albicans and other pathogenic bacteria from urethra in men and from urethra, vagina and cervix in women. Women had also cytologic examination from cervix with Papanicolau method. We have found an high incidence of urethral and vaginal asymptomatic infections, of positivity for HBV markers and some positivities of tests for syphilis. Importance of these screening examinations in the management of patients with genital warts is then discussed.