Implementation and new insights in molecular diagnostics for HIV infection

Introduction: Acquired immunodeficiency syndrome (AIDS) is a kind of acquired disease that breaks down the immune system. Human immunodeficiency virus (HIV) is the causative agent of AIDS. By the end of 2016, there were 36.7 million people living with HIV worldwide. Early diagnosis can alert infected individuals to risk behaviors in order to control HIV transmission. Infected individuals are also benefited from proper treatment and management upon early diagnosis. Thanks to the public awareness of the disease, the annual increase of new HIV infections has been slowly declining over the past decades. The advent of molecular diagnostics has allowed early detection and better management of HIV infected patients.

Areas covered: In this review, the authors summarized and discussed the current and future technologies in molecular diagnosis as well as the biomarkers developed for HIV infection.

Expert Commentary: A simple and rapid detection of viral load is important for patients and doctors to monitor HIV progression and antiretroviral treatment efficiency. In the near future, it is expected that new technologies such as digital PCR and CRISPR-based technology will play more important role in HIV detection and patient management.     

Synthesis of artemiside and its effects in combination with conventional drugs against severe murine malaria

This research describes the use of novel antimalarial combinations of the new artemisinin derivative artemiside, a 10-alkylamino artemisinin. It is a stable, highly crystalline compound that is economically prepared from dihydroartemisinin in a one-step process. Artemiside activity was more pronounced than that of any antimalarial drug in use, both in Plasmodium falciparum culture and in vivo in a murine malaria model depicting cerebral malaria (CM). In vitro high-throughput testing of artemiside combinations revealed a large number of conventional antimalarial drugs with which it was additive. Following monotherapy in mice, individual drugs reduced parasitemias to nondetectable levels. However, after a period of latency, parasites again were seen and eventually all mice became terminally ill. Treatment with individual drugs did not prevent CM in mice with recrudescent malaria, except for piperaquine at high concentrations. Even when CM was prevented, the mice developed later of severe anemia. In contrast, most of the mice treated with drug combinations survived. A combination of artemiside and mefloquine or piperaquine may confer an optimal result because of the longer half life of both conventional drugs. The use of artemiside combinations revealed a significant safety margin of the effective artemiside doses. Likewise, a combination of 1.3 mg/kg of body weight artemiside and 10 mg/kg piperaquine administered for 3 days from the seventh day postinfection was completely curative. It appears possible to increase drug concentrations in the combination therapy without reaching toxic levels. Using the drug combinations as little as 1 day before the expected death of control animals, we could prevent further parasite development and death due to CM or anemic malaria. Earlier treatment may prevent cognitive dysfunctions which might occur after recovery from CM.     

In vitro study of the anti-cancer effects of artemisone alone or in combination with other chemotherapeutic agents

Purpose

Artemisinins are now established drugs for treatment of malaria. These agents have been shown to possess impressive anti-cancer properties. We have investigated the role of artemisone (ATM), a novel derivative of artemisinin (ART) in a cancer setting both alone and in combination with established chemotherapeutic agents.

Methods

The anti-proliferative effects of ART and ATM were tested on a panel of human cancer cells in vitro using the methylthiazoletetrazolium assay, and the effect on cell cycling established by flow cytometry. Immunoblot analyses were performed to determine effects at the molecular level. Finally, ART and ATM were combined with the common anti-cancer agents oxaliplatin, gemcitabine and thalidomide.

Results

ART and ATM caused dose dependent decreases in cell number. ATM was consistently superior to ART, with IC50?s significantly lower in the former. Neither drug caused significant changes to the cell viability (%viable cells >95%), but arrested cell cycling. Blockade was either exclusively at the level of G1, or at all phases of the cell cycle, and associated with reductions in cyclin D1, CDK4 and pRb. Combination studies showed the anti-proliferative effect of ATM was often enhanced by addition of the other drugs, whilst ART exhibited antagonistic properties.

Conclusions

ART and ATM are active in cancer cell lines, with ATM displaying the greater anti-proliferative effect when used alone. ATM also enhances the effects of the above drugs, with ART being less likely to improve activities. Taken together, ATM should be thought of as the ART-derived compound next in line for further study.     

Clinical and pathological characterization of FLNC-related myofibrillar myopathy caused by founder variant c.8129G>A in Hong Kong Chinese

FLNC-related myofibrillar myopathy could manifest as autosomal dominant late-onset slowly progressive proximal muscle weakness; involvements of cardiac and/or respiratory functions are common. We describe 34 patients in nine families of FLNC-related myofibrillar myopathy in Hong Kong ethnic Chinese diagnosed over the last 12 years, in whom the same pathogenic variant c.8129G>A (p.Trp2710*) was detected. Twenty-six patients were symptomatic when diagnosed; four patients died of pneumonia and/or respiratory failure. Abnormal amorphous material or granulofilamentous masses were detected in half of the cases, with mitochondrial abnormalities noted in two-thirds. We also show by haplotype analysis the founder effect associated with this Hong Kong variant, which might have occurred 42 to 71 generations ago or around Tang and Song dynasties, and underlain a higher incidence of myofibrillar myopathy among Hong Kong Chinese. The late-onset nature and slowly progressive course of the highly penetrant condition could have significant impact on the family members, and an early diagnosis could benefit the whole family. Considering another neighboring founder variant in FLNC in German patients, we advocate development of specific therapies such as chaperone-based or antisense oligonucleotide strategies for this particular type of myopathy.     

Validation study of a portable monitoring device for identifying OSA in a symptomatic patient population

Background and objective: Obstructive sleep apnoea syndrome (OSAS) is a common disorder associated with early atherosclerosis, diabetes mellitus, ischaemic heart disease and cerebrovascular disease. The gold standard for confirming OSAS is based on an attended overnight polysomnography (PSG) in a sleep laboratory; however lack of health‐care resources creates long waiting times for patient access to this diagnostic test. This study evaluated the ability of a portable sleep‐monitoring device to identify patients in Hong Kong with suspected OSAS. Methods: Patients with symptoms of OSAS were invited to use the ARES (apnoea risk evaluation system) concurrently with an attended inpatient PSG. Several sets of AHI were generated by the ARES provider based on different oxygen desaturation criteria and surrogate parameters of arousal. The results were compared against PSG to determine the optimal sensitivity and specificity. Results: There were 141 patients who completed the study successfully. Results of AHI from the ARES study were presented in the order of different scoring criteria––4% oxygen desaturation alone, obstructive events with 3% oxygen desaturation and obstructive events with 1% desaturation plus surrogate arousal criteria. The sensitivity was 0.84 (95% confidence interval (CI): 0.77–0.90), 0.89 (95% CI: 0.89–0.94) and 0.97 (95% CI: 0.94–0.99), respectively. The specificity was 1, 1 and 0.63 (95% CI: 0.55–0.71), respectively. The receiver operating curve had an area of 0.96, 0.97 and 0.98, respectively. The kappa coefficient varied from 0.24 to 0.55 for agreement of severity between PSG and ARES. The likelihood ratio positive and the likelihood ratio negative were 2.61, infinity, infinity and 0.16, 0.11, 0.05, respectively, in the order of oxygen desaturation described earlier. Conclusions: The ARES device has reasonable sensitivity and specificity for diagnosing severe OSAS in symptomatic Chinese patients. There is moderate agreement between ARES and PSG in the diagnosis of severe disease, but less agreement in patients with mild/moderate disease.     

Current and future molecular diagnostics in colorectal cancer and colorectal adenoma

Colorectal cancer(CRC)is one of the most prevalent cancers in developed countries.On the other hand,CRC is also one of the most curable cancers if it is detected in early stages through regular colonoscopy or sigmoidoscopy.Since CRC develops slowly from precancerous lesions,early detection can reduce both the incidence and mortality of the disease.Fecal occult blood test is a widely used non-invasive screening tool for CRC.Although fecal occult blood test is simple and cost-effective in screening CRC,there is room for improvement in terms of the accuracy of the test.Genetic dysregulations have been found to play an important role in CRC development.With better understanding of the molecular basis of CRC,there is a growing expectation on the development of diagnostic tests based on more sensitive and specific molecular markers and those tests may provide a breakthrough to the limitations of current screening tests for CRC.In this review,the molecular basis of CRC development,the characteristics and applications of different non-invasive molecular biomarkers,as well as the technologies available for the detection were discussed.This review intended to provide a summary on the current and future molecular diagnostics in CRC and its pre-malignant state,colorectal adenoma.     

Absorption of the novel artemisinin derivatives artemisone and artemiside: potential application of Pheroid™ technology

Artemisinins have low aqueous solubility that results in poor and erratic absorption upon oral administration. The poor solubility and erratic absorption usually translate to low bioavailability. Artemisinin-based monotherapy and combination therapies are essential for the management and treatment of uncomplicated as well as cerebral malaria. Artemisone and artemiside are novel artemisinin derivatives that have very good antimalarial activities. Pheroid? technology is a patented drug delivery system which has the ability to entrap, transport and deliver pharmacologically active compounds. Pharmacokinetic models were constructed for artemisone and artemiside in Pheroid? vesicle formulations. The compounds were administered at a dose of 50.0mg/kg bodyweight to C57 BL/6 mice via an oral gavage tube and blood samples were collected by means of tail-bleeding. Drug concentrations in the samples were determined using an LC/MS/MS method. There was 4.57 times more artemisone in the blood when the drug was entrapped in Pheroid? vesicles in comparison to the drug only formulation (p < 0.0001). The absorption of artemiside was not dramatically enhanced by the Pheroid? delivery system.     

Caregiving burden and quality of life of pneumoconiosis caregivers in Hong Kong

Objective

Silicosis is the most common type of pneumoconiosis in Hong Kong. This study explored the clinical correlates of the caregiving burden and quality of life (QOL) among pneumoconiosis caregivers in Hong Kong.

Methods

The study sample included 112 patients with pneumoconiosis and their caregivers. Caregiving burden was measured using the Caregiving Burden Scale (CBS), and caregivers' QOL was assessed using the physical and mental components (PCS and MCS, respectively) of the Medical Outcomes Short Form-36. Pearson's correlation coefficient and Spearman's ρ were calculated to examine the relationship between CBS, PCS, and MCS scores and caregivers' and patients' sociodemographic variables. Stepwise regression analyses were performed to determine the independent correlates of CBS, PCS, and MCS scores.

Results

Caregiving burden was correlated with certain patient characteristics (duration of disease, severity of dyspnea, exercise tolerance, depressive symptoms, daily functioning, and community living skills) and with caregivers' variables (depressive symptoms and availability of family support). Regression analysis showed that patients' daily functioning (β = −.345), caregivers' depressive symptoms (β = .509), and the availability of family support (β = .240) were independent correlates of caregiving burden, explaining 45% of the variance. The independent correlates of PCS included patients' severity of coexisting diseases (β = −.179) and caregivers' depressive symptoms (β = −.521). Both patients' (β = −.155) and caregivers' (β = −.633) depressive symptoms and patients' severity of dyspnea (β = −.183) were independent correlates of the MCS.

Conclusion

Caring for pneumoconiosis patients entails a significant caregiving burden for caregivers, and adversely affects their QOL. Caregivers' depressive symptoms are related to both their caregiving burden and QOL.     

Trabecular bone status in ultradistal tibia under habitual gait loading: a pQCT study in postmenopausal women.

This study investigated regional volumetric trabecular bone mineral density (tBMD) and bone area at the ultradistal tibia in Chinese women using peripheral quantitative computed tomography. Fifty-six postmenopausal women aged 47-62 yr participated in BMD measurements at baseline and 22 of them were followed at both 1-yr and 3-yr follow-up scans. Regional baseline tBMD, rate of annual bone loss, and trabecular bone area were determined. Baseline measurements showed that the tBMD of both the posterior (252.9+/-63.4 mg/cm(3)) and medial (226.6+/-68.9 mg/cm(3)) regions was significantly higher than that of the anterior (126.3+/-61.9 mg/cm(3)) and lateral regions (149.8+/-50.6 mg/cm(3)), respectively (p<0.001). Both the 1-yr and 3-yr follow-up measurements showed that there was significant physiological annual tBMD loss on an average of 1.61%, at the four regions. Inter-slice regional tBMD and trabecular bone area measurements demonstrated a significant linear decrease from the distal to proximal aspects (p<0.001). Findings suggest that dynamic compressive loading during the heel strike and the body weight vector shifting toward the medial aspect during the stance phase in a normal gait might account for the regional tBMD differences. Increased tBMD and bone area toward the distal tibial endplate may adapt to withstand the axial impact loading. However, the low-impact weight-bearing nature of a normal gait may not be osteogenic to prevent regional bone loss. An exercise program specific to the women at risk should be contemplated.     

Children with islet autoimmunity and enterovirus infection demonstrate a distinct cytokine profile

Cytokines are upregulated in prediabetes, but their relationship with Enterovirus (EV) infection and development of islet autoimmunity is unknown. Cytokines (n = 65) were measured using Luminex xMAP technology in a nested case-control study of 67 children with a first-degree relative with type 1 diabetes: 27 with islet autoantibodies (Ab(+)) and 40 age-matched persistently autoantibody negative (Ab(-)) control subjects. Of 74 samples, 37 (50%) were EV-PCR(+) in plasma and/or stool (EV(+)) and the remainder were negative for EV and other viruses (EV(-)). Fifteen cytokines, chemokines, and growth factors were elevated (P ≤ 0.01) in Ab(+) versus Ab(-) children (interleukin [IL]-1β, IL-5, IL-7, IL-12(p70), IL-16, IL-17, IL-20, IL-21, IL-28A, tumor necrosis factor-α, chemokine C-C motif ligand [CCL]13, CCL26, chemokine C-X-C motif ligand 5, granulocyte-macrophage colony-stimulating factor, and thrombopoietin); most have proinflammatory effects. In EV(+) versus EV(-) children, IL-10 was higher (P = 0.005), while IL-21 was lower (P = 0.008). Cytokine levels did not differ between Ab(+)EV(+) and Ab(+)EV(-) children. Heat maps demonstrated clustering of some proinflammatory cytokines in Ab(+) children, suggesting they are coordinately regulated. In conclusion, children with islet autoimmunity demonstrate higher levels of multiple cytokines, consistent with an active inflammatory process in the prediabetic state, which is unrelated to coincident EV infection. Apart from differences in IL-10 and IL-21, EV infection was not associated with a specific cytokine profile.