Serine protease inhibitors LmSPN2 and LmSPN3 co-regulate embryonic diapause in Locusta migratoria manilensis(Meyen)via the Toll pathway

Female adults of the migratory locust,Locusta migratoria manilensis(Meyen),can sense seasonal photoperiod changes,which induces embryonic diapause as a key strategy to overwinter.Serine protease inhibitor genes(SPNs)were thought to play key roles during diapause,while few SPNs were functionally characterized.LmSPN2 was one of those genes differentially expressed between diapause and non-diapause eggs;however,its biological function remained to be explored.So,we conducted RNAi knockdown of LmSPN2,resulting in a significant decrease of the egg diapause rate by 29.7%.Using yeast two-hybrid assays,co-immunoprecipitation,and pull-down methods,we found an interaction between LmSPN2 and LmSPN3,which was proved to be mediated by a glutamate(E331)binding site of LmSPN2.RNAi knockdown of LmSPN3 resulted in a significant increase in diapause rate by 14.6%,indicating an inverse function of LmSPN2 and LmSPN3 on diapause regulation.Double knockdown of two SPN genes resulted in a 26.4%reduction in diapause rate,indicating that LmSPN2 was the dominant regulatory signal.Moreover,we found four Toll pathway genes(easter,spätzle,pelle,and dorsal)upregulated significantly after the knockdown of LmSPN2 while downregulated after the knockdown of LmSPN3.Therefore,we speculate that two SPNs regulate diapause through the Toll pathway.Our results indicated that LmSPN2 positively regulates locust egg entry into diapause,while LmSPN3 is a negative regulator of embryonic commitment to diapause.Their interaction is mediated by the binding site of E331 and influences egg diapause through the Toll pathway.This mechanistic understanding of diapause regulation expands our understanding of insect developmental regulation and provides functional targets for developing locust management strategies.

Potato protease inhibitors,a functional food material with antioxidant and anticancer potential

Potato protease inhibitors(PPIs),as the main component of potato protein isolate,have good safety,nutrition and great market potential.The antioxidant and anticancer properties of PPIs were evaluated with cellbased biological assays.The results showed that when the concentration of PPIs was 5 mg/mL,the peroxyl radical scavenging value was(2119±204)mg VCE/100 g,and the cellular antioxidant activity values were(45.83±3.5)(no PBS wash)and(33.25±4.4)μmol QE/100 g(PBS wash).Cells pretreated with PPIs could significantly prevent the oxidative damage induced by H_(2)O_(2),inhibit the morphological changes of cells and maintain the integrity.Furthermore,PPIs had selective anti-proliferative effects on GIST882 cells(IC50=(10.53±3.87)mg/mL)and demonstrated potent inhibition of the growth,migration and invasion of cancer cells.These findings provide a scientific basis for PPIs as promising candidates for functional foods to aid in the prevention of oxidative damage and cancer.

Hepatitis C virus protease inhibitor-resistance mutations: Our experience and review

Direct-acting antiviral agents(DAAs)for hepatitis C virus(HCV)infection are one of the major advances in its medical treatment.The HCV protease inhibitors boceprevir and telaprevir were the first approved DAAs in the United States,Europe,and Japan.When combined with peginterferon plus ribavirin,these agents increase sustained virologic response rates to70%-80%in treatment-na?ve patients and previoustreatment relapsers with chronic HCV genotype 1 infection.Without peginterferon plus ribavirin,DAA monotherapies increased DAA-resistance mutations.Several new DAAs for HCV are now in clinical development and are likely to be approved in the near future.However,it has been reported that the use of these drugs also led to the emergence of DAA-resistance mutations in certain cases.Furthermore,these mutations exhibit cross-resistance to multiple drugs.The prevalence of DAA-resistance mutations in HCV-infected patients who were not treated with DAAs is unknown,and it is as yet uncertain whether such variants are sensitive to DAAs.We performed a population sequence analysis to assess the frequency of such variants in the sera of HCV genotype 1-infected patients not treated with HCV protease inhibitors.Here,we reviewed the literature on resistance variants of HCV protease inhibitors in treatment na?ve patients with chronic HCV genotype 1,as well as our experience.

NS3 protease inhibitors for treatment of chronic hepatitis C: Efficacy and safety

A new treatment paradigm for hepatitis C is that the treatment must include an existing direct-acting antiviral agent, namely, a protease inhibitor(PI) combined with PEGylated interferon-a and ribavirin. The currently mar-keted PIs and PIs in clinical trials have different mecha-nisms of action. The development of new PIs aims for an improved safety profile and higher effectiveness. This article reviews NS3/4A protease inhibitors, focusing on major criteria such as their effectiveness and safety. Specific attention is paid to dosing regimens and adverse event profiles of PIs administered in clinical settings.

Comparative Study on the Protease Inhibitors from Fish Eggs

The protease inhibitor was purified from five different fish eggs. The molecular weights of Pacific herring, chumsalmon, pond smelt, glassfish, and Alaska pollock egg protease inhibitors were 120, 89, 84.5, 17, and 16.8 kDa, respective-ly. The specific inhibitory activity of glassfish egg protease inhibitor was the highest followed by those of Pacific herring andAlaska pollock in order. The specific inhibitory activity and purity of glassfish egg protease inhibitor were 19.70 U mg -1 pro-tein and 164.70 folds of purification, respectively. Glassfish egg protease inhibitor was reasonably stable at 50 - 65 °C and pH 8,which was more stable at high temperature and pH than protease inhibitors from the other fish species. Glassfish egg pro-tease inhibitor was noncompetitive with inhibitor constant (Ki) of 4.44 nmol L-1.

HIV aspartyl protease inhibitors as promising compounds against Candida albicans

Cells of Candida albicans(C.albicans) can invade humans and may lead to mucosal and skin infections or to deep-seated my coses of almost all inner organs,especially in immunocompromised patients.In this context,both the host immune status and the ability of C.albicans to modulate the expression of its virulence factors are relevant aspects that drive the candidal susceptibility or resistance;in this last case,culminating in the establishment of successful infection knownas candidiasis.C.albicans possesses a potent arma-mentarium consisting of several virulence moleculesthat help the fungal cells to escape of the host immuneresponses.There is no doubt that the secretion of aspartyl-type proteases,designated as Saps,are one of the major virulence attributes produced by C.albicans cells,since these hydrolytic enzymes participate in a wide range of fungal physiological processes as well as in different facets of the fungal-host interactions.For these reasons,Saps clearly hold promise as new potential drug targets.Corroborating this hypothesis,the introduction of new anti-human immunodeficiency virus drugs of the as party l protease inhibitor-type(HIV PIs) have emerged as new agents for the inhibition of Saps.The introduction of HIV PIs has revolutionized the treatment of HIV disease,reducing opportunistic infections,especially candidiasis.The attenuation of candidal infections in HIV-infected individuals might not solely have resulted from improved immunological status,but also as a result of direct inhibition of C.albicans Saps.In this article,we review updates on the beneficial effects of HIV PIs against the human fungal pathogen C.albicans,focusing on the effects of these compounds on Sap activity,growth behavior,morphological architecture,cellular differentiation,fungal adhesion to animal cells and abiotic materials,modulation of virulence factors,experimental candidiasis infection,and their synergistic actions with classical antifungal agents.

Molecular mechanism of modulating the liquefaction epididymal protease inhibitor of human semen

Aim： To study the molecular mechanism of epididymal protease inhibitor （Eppin） modulating the process of prostate specific antigen （PSA） digesting semenogelin （Sg）. Methods： Human Sg cDNA （nucleotides 82-849） and Eppin cDNA （nucleotides 70-423） were generated by polymerase chain reaction （PCR） and cloned into pET-100D/TOPO. Recombinant Eppin and Sg （rEppin and rSg） were produced by BL21 （DE3）. The association of Eppin with Sg was studied by far-western immunoblot and radioautography. In vitro the digestion of rSg by PSA in the presence or absence of rEppin was studied. The effect of anti-Q20E （N-terminal） and C-terminal of Eppin on Eppin-Sg binding was monitored. Results： Eppin binds Sg on the surface of human spermatozoa with the C-terminal of Eppin （amino acids 75-133）. rSg was digested with PSA and many low molecular weight fragments were produced. When rEppin is bound to rSg, then digested by PSA, incomplete digestion and a 15-kDa fragment results. Antibody binding to the N-terminal of rEppin did not affect rSg digestion. Addition of antibodies to the C-terminal of rEppin inhibited the modulating effect of rEppin. Conclusion： Eppin protects a 15-kDa fragment of rSg from hydrolysis by PSA.

Association between calcium sensing receptor gene polymorphisms and chronic pancreatitis in a US population:Role of serine protease inhibitor Kazal 1type and alcohol

AIM: To test the hypothesis that calcium sensing receptor (CASR) polymorphisms are associated with chronic pancreatitis (CP), and to determine whether serine protease inhibitor Kazal 1type (SPINK1) N34S oralcohol are necessary co-factors in its etiology. METHODS: Initially, 115 subjects with pancreatitis and 66 controls were evaluated, of whom 57 patients and 21 controls were predetermined to carry the high-risk SPINK1 N34S polymorphism. We sequenced CASR gene exons 2, 3, 4, 5 and 7, areas containing the majority of reported polymorphisms and novel mutations. Based on the initial results, we added 223 patients and 239 controls to analyze three common nonsynonymous single nucleotide polymorphisms (SNPs) in exon 7 (A986S, R990G, and Q1011E). RESULTS: The CASR exon 7 R990G polymorphism was signifi cantly associated with CP (OR, 2.01; 95% CI, 1.12-3.59; P = 0.015). The association between CASR R990G and CP was stronger in subjects who reported moderate or heavy alcohol consumption (OR, 3.12; 95% CI, 1.14-9.13; P = 0.018). There was no association between the various CASR genotypes and SPINK1 N34S in pancreatitis. None of the novel CASR polymorphisms reported from Germany and India was detected. CONCLUSION: Our United States-based study confirmed an association of CASR and CP and for the first time demonstrated that CASR R990G is a signifi cant risk factor for CP. We also conclude that the risk of CP with CASR R990G is increased in subjects with moderate to heavy alcohol consumption.

Effects of inhibitors on the protease profiles and degradation of activated Cry toxins in larval midgut juices of Cnaphalocrocis medinalis (Lepidoptera:Pyralidae)

Midgut juice plays an important role in food digestion and detoxification in insects.In order to understand the potential of midgut juice of Cnaphalocrocis medinalis(Guenée)to degrade Bt proteins,the enzymatic activity of midgut juice and its degradation of Bt proteins(Cry2A,Cry1C,Cry1Aa,and Cry1Ac)were evaluated in this study through protease inhibitor treatments.The activities of total protease in midgut juices were significantly inhibited by phenylmethylsulfonyl fluoride(PMSF),tosyl-L-lysine chloromethyl ketone(TLCK),pepstatin A and leupeptin.The enzymatic activity of chymotrypsin was significantly inhibited by PMSF,and enzymatic activity of trypsin was significantly inhibited by ethylenediaminetetraacetic acid(EDTA),PMSF,tosyl phenylalanine chloromethyl ketone(TPCK),TLCK and trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane(E-64).EDTA could significantly inhibit the degradation of Cry2A by C.medinalis.EDTA,PMSF,TPCK,and TLCK could inhibit the degradation of Cry1C and Cry1Aa.EDTA,PMSF,TPCK,TLCK,and E-64 could inhibit the degradation of Cry1Ac.Our results indicated that some protease inhibitors hindered various enzymatic activities in the larval midgut of C.medinalis,which may reduce the insect’s ability to degrade Bt toxins.These findings may aid the application of protease inhibitors in the management of this insect pest in the future.

Emerging potential of ubiquitin-specific proteases and ubiquitinspecific proteases inhibitors in breast cancer treatment

Breast cancer is the most frequently diagnosed cancer in women,accounting for 30%of new diagnosing female cancers.Emerging evidence suggests that ubiquitin and ubiquitination played a role in a number of breast cancer etiology and progression processes.As the primary deubiquitinases in the family,ubiquitin-specific peptidases(USPs)are thought to represent potential therapeutic targets.The role of ubiquitin and ubiquitination in breast cancer,as well as the classification and involvement of USPs are discussed in this review,such as USP1,USP4,USP7,USP9X,USP14,USP18,USP20,USP22,USP25,USP37,and USP39.The reported USPs inhibitors investigated in breast cancer were also summarized,along with the signaling pathways involved in the investigation and its study phase.Despite no USP inhibitor has yet been approved for clinical use,the biological efficacy indicated their potential in breast cancer treatment.With the improvements in phenotypic discovery,we will know more about USPs and USPs inhibitors,developing more potent and selective clinical candidates for breast cancer.

Molecular mechanism of epididymal protease inhibitor modulating the liquafication of human semen

To study the molecular mechanism of epididymal protease inhibitor （Eppin） modulating the liquafication of semen. Methods： Human semenogelin cDNA （nucleotides 82-849） and Eppin cDNA （nucleotides 70-423） were generated by PCR and cloned into pET-100D/TOPO.Recombinant Eppin and Sg were produced by BL21 （DE3）. The association of Eppin with Sg was studied by far-western and radioautography.In vitro the digestion of Sg by PSA in the presence or absence of recombinant Eppin was studied. The effect of anti-Q20E （N-terminal） and C-terminal of Eppin on Eppin-Sg binding was monitored. Results： Eppin binds Sg on the surface of human spermatozoa with C-terminal Eppin （aa75-133）.Recombinant Sg was digested with PSA ,many low molecular weight fragments were produced, when Eppin is bound to Sg ,then digested by PSA ,producing incomplete digestion and a 14.5-14.8 kDa fragmen. Antibody binding to the N-terminal of Eppin did not affect Sg digestion. Addition of antibodies to the C-terminal of Eppin inhibited the modulating effects of Eppin. Conclusion： Eppin modulates the digestion activity of PSA through binding Sg.The active site locates at C-terminal.

A Study on Heat Resistance and Initial Characterization of Protease Inhibitors in Porcine Colostrum

Porcine colostrum was separated into the acid soluble fraction (SF) and casein fraction (CF) by acidifying followed by centrifuge. SF was further separated by liquid chromatography and anisotropic membrane filtration. Capacities of the SF or CF of porcine colostrum, to inhibit trypsin and chymotrypsin activity and to inhibit the epidermal growth factor (EGF) degradation in pig small intestinal contents, were determined under different heat treatments. The study showed that trypsin inhibitors in porcine colostrum survived heat treatments of 100℃ water bath for up to 10 min, but exposure to boiling water bath for 30 min significantly decreased the inhibitory activity. Compared with the trypsin inhibitors, the chymotrypsin inhibitors were more heat sensitive. SF was more heat sensitive than CF. Separation of the SF of porcine colostrum by liquid chromatography and anisotropic membrane filtration revealed that the porcine colostrum protease inhibitors, those had the capacity to inhibit the trypsin chymotrypsin activity and enhanced the stability of EGF in the gastrointestinal(GI) lumen of weaned pigs, existed mainly in SF, milk derived, were a group of heat labile small proteins with molecular weight of 10 00050 000.

A Study on the Inhibitory Potency of Protease Inhibitorsin Porcine Colostrum

Porcine colostrum and milk were separated into the acid-soluble fraction(SF)and casein fraction(CF)by centrifuge. Trypsin and chymotrypsin inhibitory capacity in porcine colostrum, milk and their components were determined by incubating bovine trypsin or chymotrypsin in a medium. The inhibition of insulin-like growth factor I(IGF-I)and epidermal growth factor(EGF)degradation in pig small intestinal contents by porcine colostrum was measured by incubating iodinated IGF-I or EGF. Degradation of labeled IGF-I or EGF was determined by monitoring the generation of radioactivity soluble in 30% trichloroacetic acid(TCA). The results showed that porcine colostrum had high levels of trypsin and chymotrypsin inhibitory activity and increased the stability of IGF-I and EGF in pig intestinal contents. The SF was higher in inhibitory potency than CF. The present study revealed that the protease inhibitors in porcine colostrum, milk-derived and colostrum-specific, existed mainly in SF.

Protease Inhibitors in Wild Relatives of Pigeonpea against the Cotton Bollworm/Legume Pod Borer, <i>Helicoverpa armigera</i>

Cotton bollworm/legume pod borer, Helicoverpa armigera is one of the most damaging pests worldwide. Be-cause of the difficulties associated with chemical control of this pest, emphasis has been placed on developing transgenic plants with resistance to H. armigera. Since toxin genes from the bacterium, Bacillus thuringien-sis (Bt) have been deployed on a large scale, there is need to scout for alternate genes which could be deployed alone or in combination with the Bt genes for pest management. Therefore, we evaluated the wild relatives of pigeonpea, which have shown high levels of resistance to this pest, for the protease inhibitors (PIs) under in vivo and in vitro inhibitions. Accessions belonging to Cajanus albicans, C. cajanifolius, C. sericeus, Flemingia bracteata, and Rhynchosia bracteata showed complete inhibition of H. armigera gut proteinases (HaGPs). Some of the C. scarabaeoides accessions (ICPW 116, 152, 278 and 280) exhibited partial inhibition at low concentrations of the PIs. All accessions of wild relatives of pigeonpea showed high to moderate level of inhibition at pH 7.8. Cultivated pigeonpea, ICPL 87 exhibited monomorphism in terms of trypsin inhibitor (TI) and chymotrypsin inhibitor (CTI) isoforms, contrary to the diverse inhibitory profiles of wild pigeonpeas. Cajanus albicans, C. platycarpus, C. scarabaeoides, and R. bracteata showed more number of TI and CTI bands than the cultivated pigeonpea. Protease inhibitor isoforms of wild relatives of pigeonpea showed significant variation in number, band pattern, and protein specificities towards trypsin, chymotrypsin, and H. armigera gut proteinases (HaGPs) as compared to the cultivated pigeonpea. The PIs from the wild relatives of pigeonpea showed considerable potential against the HaGPs, and could be considered as potential candidates for use in genetic transformation of crops for pest management, including H. armigera.

HIV protease inhibitors sensitize human head and neck carcinoma cells to radiation by activating endoplasmic reticulum stress

Aim Head and neck cancers are the eighth most common cancer worldwide. Despite significant ad- vances in the delivery of treatment and surgical reconstruction, the mortality rates for this disease have not improved in the past 4 decades. Our previous study has shown that HIV protease inhibitors （HIV PIs） induce cell apoptosis via activating endoplasmic reticulum （ER） stress. It also has been reported that a few HIV PIs are able to radio- sensitize tumor cells. However, the underlying cellular mechanisms remain to be identified. The aim of this study was to examine whether HIV PIs activate the ER stress response and sensitize human head and neck carcinoma cells to radiation. Methods Human SQ20B and Fadu cells and the most commonly used HIV PIs, lopinavir and ritona- vir, were used in this study. The mRNA and protein levels of ER stress-related genes （ CHOP, ATF4, XBP-1, and GRP78 ） were detected by real time RT-PCR and Western blot, respectively. Cell viability and apoptosis were ana- lyzed using Cellometer Vision CBA. After treatment with HIV PIs, cells were irradiated at a dose of 2G or 4G. Col- onies were stained and counted 10 days after irradiation. Results HIV PIs significantly induced activation of ER stress and apoptosis. Treatment of HIV PIs inhibited Akt phosphorylation, induced cell cycle arrest in G1 phase and increased tumor cell sensitivity to irradiation-induced cell death. Conclusion HIV PIs sensitize human head and neck carcinoma cells to radiation by activating ER stress.

Effectiveness and safety of first-generation protease inhibitors in clinical practice:Hepatitis C virus patients with advanced fibrosis

AIM:To evaluates the effectiveness and safety of the first generation,NS3/4A protease inhibitors(PIs) in clinical practice against chronic C virus,especially in patients with advanced fibrosis. METHODS:Prospective study and non-experimental analysis of a multicentre cohort of 38 Spanish hospitals that includes patients with chronic hepatitis C genotype 1,treatment-na?ve(TN) or treatment-experienced(TE),who underwent triple therapy with the first generation NS3/4A protease inhibitors,boceprevir(BOC) and telaprevir(TVR),in combination with pegylated interferon and ribavirin. The patients were treatment in routine practice settings. Data on the study population and on adverse clinical and virologic effects were compiled during the treatment period and during follow up.RESULTS:One thousand and fifty seven patients were included,405(38%) were treated with BOC and 652(62%) with TVR. Of this total,30%(n = 319) were TN and the remaining were TE:28%(n = 298) relapsers,12%(n = 123) partial responders(PR),25%(n = 260) null-responders(NR) and for 5%(n = 57) with prior response unknown. The rate of sustained virologic response(SVR) by intention-to-treatment(ITT) was greater in those treated with TVR(65%) than in those treated with BOC(52%)(P < 0.0001),whereas by modified intention-to-treatment(m ITT) no were found significant differences. By degree of fibrosis,56% of patients were F4 and the highest SVR rates were recorded in the non-F4 patients,both TN and TE. In the analysis by groups,the TN patients treated with TVR by ITT showed a higher SVR(P = 0.005). However,by m ITT there were no significant differences between BOC and TVR. In the multivariate analysis by m ITT,the significant SVR factors were relapsers,IL28 B CC and non-F4; the type of treatment(BOC or TVR) was not significant. The lowest SVR values were presented by the F4-NR patients,treated with BOC(46%) or with TVR(45%). 28% of the patients interrupted the treatment,mainly by non-viral response(51%):this outcome was more frequent in the TE than in the TN patients(57% vs 40%,P = 0.01). With respect to severe haematological disorders,neutropaenia was more likely to affect the patients treated with BOC(33% vs 20%,P ≤ 0.0001),and thrombocytopaenia and anaemia,the F4 patients(P = 0.000,P = 0.025,respectively). CONCLUSION:In a real clinical practice setting with a high proportion of patients with advanced fibrosis,effectiveness of first-generation PIs was high except for NR patients,with similar SVR rates being achieved by BOC and TVR.

Synthesis of N1-Substituted-3-aryl-4-alkyl-4, 5-dihydro-1H-1-pyra-zolethiocarboxamide as Novel Small Molecule Inhibitors of Cysteine Protease of T.cruzi

A series of N1-substituted-3-aryl-4-alkyl-4, 5-dihydro-1H-1-pyrazolethiocarboxamide were prepared from the Mannich bases of aryl ketones in good yields. Some derivatives were found to be active against the cysteine protease of T.cruzi..

Effect of protease inhibitor from Agaricus bisporus on glucose uptake and oxidative stress in 3T3-L1 adipocytes

Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.

Do Protease Inhibitors Increase Preterm Births in Human Immunodeficiency Virus-Infected Patients?

Objective: To compare preterm delivery (PTD) rates in HIV-infected patients on a protease inhibitor (PI)-based and a PI-sparing regimen. Study Design: This is a retrospective review of records of HIV-infected pregnant women between 2000 and 2007 at University Hospital, Newark, NJ. Patients were grouped according to PI exposure during pregnancy. Rates of preterm birth were compared, and the analysis was performed irrespectively of the etiology or indication of the preterm birth. Multivariate analysis including substance use, PI use, initial CD4 count, and history of PTD was performed. Results: There were 129 pregnant women in the PI group and 59 in the PI-sparing group. The PTD rate did not differ between the PI group and PI-sparing group (27.9% vs 25.4%, P = 0.72). 28.6% of those who delivered preterm had a previous PTD compared to 8.4% of those who delivered at term (P = 0.0019). Patients who delivered preterm had a higher rate of substance use (37.3% vs 19.7%, P = 0.0128). In the multivariate analysis, only history of PTD was significant (P = 0.018). Conclusion: Contrary to other studies, PIs were not associated with PTD. Other known risk factors of PTD, specifically past PTD and substance use, should be considered and targeted for risk reduction during pregnancy.

Macromolecular inhibitors of malarial cysteine proteases —An invited review

There are evidences indicating that cysteine proteases play an essential role in malaria parasites;therefore, an obvious area of investigation is the inhibition of these enzymes to treat malaria. Small cysteine protease inhibitors of malaria are well studied, but macromolecular nature of inhibitor is a new field to explore. In malarial cysteine proteases, there are macromolecular endogenous inhibitors playing important roles in regulation of the cysteine protease activity of parasite and host. Recent studies suggested that there are known and characterized endogenous inhibitors like falstatin present in P. falciparum, PbICP (inhibitor of cysteine protease in P. berghei), PyICP (inhibitor of cysteine protease in P. yoelli), and other macromolecular inhibitors which are the prodomain of enzyme itself regulating the activity of the mature enzyme. All the known macromolecular endogenous inhibitors are using specific loop-like structure to interact with malarial cysteine proteases. The majority of macromolecular inhibitors are competitive in nature, and block access to the active site of their target protease, but do not bind in a strictly substrate-like manner. They rather interact with the protease subsites and catalytic residues in a non-catalytically competent manner. In future, designing inhibitors based on these protein-protein interactions will be a new approach in the field of malaria. Since macromolecular inhibitors can gain potency through the burial of a large surface area and specificity through contacts with secondary binding sites critical for inhibition, and could be less prone to drug resistant mutation.